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Primary Antibodies

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To determine the titer, an ELISA was performed using a serial dilution of the antibody directed against human MBD4 (bs-53050R) in antigen-coated wells. The wells were coated with peptides. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:15,000.
(Left) U2OS cells were transfected with an expression vector for TY1-tagged MBD4. The presence of TY1-MBD4 in the cell lysates was demonstrated by western blot analysis with the antibody against the TY1-tag (lane 1) and with the antibody against MBD4 (bs-53050R) (lane 2), diluted 1:2,000 in TBST containing 3% milk powder. (Right) Western blot was performed on nuclear extracts from the U937 human leukemic monocyte lymphoma cell line (40 \u03bcg) with the antibody against MBD4 (bs-53050R), diluted 1:2,000 in TBST containing 3% milk powder.

Catalog #1: bs-53050R

Applications: WB, ELISA

Reactivity: Human

Conjugate: Unconjugated

Catalog #1: bs-53056R

Applications: ELISA

Reactivity: Mouse

Conjugate: Unconjugated

To determine the titer of the antibody, an ELISA was performed using a serial dilution MYH11 Polyclonal Antibody (bs-53091R). The plates were coated with the peptides. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:1,900.
ChIP assays were performed using ME-1 cells, MYH11 Polyclonal Antibody (bs-53091R) and optimized primer pairs for qPCR. Sheared chromatin from 1.5 million cells and 5 \u03bcl of antibody were used per ChIP experiment. QPCR was performed using primers specific for the genes indicated. The figure shows the relative occupancy, calculated as the ratio and control\/background for which the MYOG gene was used.
The IP\u2019d DNA from 6 ChIP\u2019s was pooled and subsequently analyzed on an Illumina HiSeq 2000. Library preparation, cluster generation, and sequencing were performed according to the manufacturer\u2019s instructions. The 50 bp tags were aligned to the human genome using the BWA algorithm. The figure shows the signal in 4 genomic regions surrounding the AXIN1, FUT7, BCL3 and RAD50 positive control genes.

Catalog #1: bs-53091R

Applications: WB, ELISA, ChIP-seq

Reactivity: Human

Conjugate: Unconjugated

Catalog #1: bs-53085R

Applications: WB, ELISA

Reactivity: Human, Mouse

Conjugate: Unconjugated

Human embryonic kidney cells (293T) were transfected with an ER-tagged PML\/RAR fusion protein. Nuclear extracts (50 \u03bcg) from control (lane 1) and from transfected cells (lane 2) were analyzed by Western blot using PML Polyclonal Antibody (Cat. No. bs-53134R). The antibody was diluted 1:750 in TBS-Tween containing 5% milk powder. The smaller band of approximately 60 kDa might represent the endogenous PML protein.
To determine the titer of the antibody, an ELISA was performed using a serial dilution of PML Polyclonal Antibody (bs-53134R), crude serum and Flow-Through in antigen-coated wells. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:6,000.

Catalog #1: bs-53134R

Applications: WB, ELISA

Reactivity: Human

Conjugate: Unconjugated

Catalog #1: bs-53084R

Applications: WB, ELISA

Reactivity: Mouse

Conjugate: Unconjugated

Western blot was performed on whole cell lysates from mouse fibroblasts (NIH3T3) and embryonic stem cells (E14Tg2a) with the DNMT1 antibody (bs-53048R), diluted 1:500 in BSA\/ PBS-Tween. The molecular weight marker (in kDa) is shown on the left; the location of the protein of interest (expected size: 183 kDa) is indicated on the right.
To determine the titer, an ELISA was performed using a serial dilution of the DNMT1 antibody (bs-53048R). The plates were coated with peptides. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:9,600.

Catalog #1: bs-53048R

Applications: WB, ELISA

Reactivity: Mouse

Conjugate: Unconjugated

Catalog #1: bs-53078R

Applications: ELISA

Reactivity: Human, Mouse

Conjugate: Unconjugated

To determine the titer, an ELISA was performed using a serial dilution of the antibody directed against H4K8ac Polyclonal Antibody (bs-53063R). The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:17,500.
A Dot Blot analysis was performed to test the cross-reactivity of \r\nH4K8ac Polyclonal Antibody (bs-53063R) with peptides containing other modifications of histone H4 and H3 and an unmodified histone H4 sequence. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. The figure shows a high specificity of the antibody for the modification of interest.
Histone extracts of HeLa cells (15 \u03bcg) were analyzed by Western blot using the antibody against H4K8ac (bs-53063R) diluted 1:250 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the left; the marker (in kDa) is shown on the right. Lane 2 shows the result of the Western analysis with the antibody; lane 1 shows the same analysis after incubation of the antibody with 750 pmol blocking peptide for 1 hour at room temperature.
ChIP assays were performed using HeLa cells treated with colcemid, H4K8ac Polyclonal Antibody (bs-53063R) and optimized PCR primer sets for qPCR. ChIP was performed using sheared chromatin from 10,000 cells. A titration of the antibody consisting of 0.5, 1.5, and 5 \u03bcl per ChIP experiment was analyzed. Additionally, ChIP was performed after incubation of the antibody with 5 nmol blocking peptide for 1 hour at room temperature. IgG (5 \u03bcg\/IP) was used as negative IP control. QPCR was performed with primers for the promoter of the active genes ALDOA and MYOD. The figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

Catalog #1: bs-53063R

Applications: WB, ELISA, dot-blot

Reactivity: Human

Conjugate: Unconjugated

Histone extracts of HeLa cells (15 \u03bcg) were analysed by Western blot using the H4K20me1 Polyclonal Antibody (bs-53035R) diluted 1:750 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.
To check the specificity of H4K20me1 Polyclonal Antibody (bs-53035R) at 1:20,000 dilution, a Dot Blot was performed with peptides containing different modifications of histone H4 and the unmodified sequence. 0.2 to 100 pmol concentration of peptides were spotted on to a membrane. Experiment shows specificity of the antibody for the modification of interest.
To determine the titer, an ELISA was performed using a serial dilution of the H4K20me1 Polyclonal Antibody (bs-53035R), to detect peptide containing histone modification of interest. By plotting the absorbance against antibody dilutions, titer of the antibody was estimated to be 1:8,000.
ChIP were performed using U2OS cells using H4K20me1 Polyclonal Antibody (bs-53035R). Chromatin was sheared from 1.6 million cells. A titration of the antibody consisting of 2, 5, 10 and 15\u03bcl per ChIP experiment was analyzed. IgG (5\u03bcg\/IP) was used as negative IP control. Quantitative PCR was performed with primers for the GAPDH promoter and for the coding region of the myogenic differentiation gene (MYOD) which is inactive at normal conditions. This figure shows the recovery, expressed as a % of input.

Catalog #1: bs-53035R

Applications: WB, ELISA, dot-blot

Reactivity: Human, Others

Conjugate: Unconjugated

Western blot was performed on whole cell extracts (25 \u03bcg, lane 1) and histone extracts (15 \u03bcg, lane 2) from HeLa cells, and on 1 \u03bcg of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using H4pan Polyclonal Antibody(bs-53138R). The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.
HeLa cells were stained with H4pan Polyclonal Antibody (bs-53138R) and with DAPI. Cells were fixed with 4% formaldehyde for 10\u2019 and blocked with PBS\/TX-100 containing 1% BSA. The cells were immunofluorescently labeled with the H4 antibody (middle) diluted 1:1,000 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The left panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.
To determine the titer of the antibody, an ELISA was performed using a serial dilution of H4pan Polyclonal Antibody (bs-53138R) in antigen coated wells. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:3,000.
ChIP assays were performed using HeLa cells, H4pan Polyclonal Antibody (bs-53138R) and optimized PCR primer pairs for qPCR. ChIP was performed using sheared chromatin from 1 million cells on the IP-Star Compact automated system. A titration consisting of 0.5, 1, 2 and 5 \u03bcg of antibody per ChIP experiment was analyzed. IgG (1 \u03bcg\/IP) was used as a negative IP control. Quantitative PCR was performed with primers specific for the promoters of the active GAPDH and c-fos genes, and for the inactive MYOD1 gene and the Sat2 satellite repeat. Figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

Catalog #1: bs-53138R

Applications: WB, ELISA, IF(ICC)

Reactivity: Human

Conjugate: Unconjugated

Mouse NIH3T3 cells were stained with Bioss antibody against H4K5,8,12,16ac (cat. No. bs-53098R) and with DAPI. Cells were fixed with 4% formaldehyde for 10\u2019 and blocked with PBS\/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labeled with the H4K5,8,12,16ac antibody (left) diluted 1:500 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.
To test the cross-reactivity of the Bioss antibody against H4K5,8,12,16ac (cat. No. bs-53098R), a Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H4. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:20,000. The figure shows a high specificity of the antibody for the modification of interest.
To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Bioss antibody directed against H4K5,8,12,16ac (cat. No. bs-53098R) in antigen-coated wells. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:21,200.
ChIP was performed with 2 \u03bcg of the Bioss antibody against H4K5,8,12,16ac (cat. No. bs-53098R) on sheared chromatin from 1 million HeLa cells using a ChIP-seq kit as described above. The IP\u2019d DNA was subsequently analyzed on an Illumina Genome Analyzer. Library preparation, cluster generation, and sequencing were performed according to the manufacturer\u2019s instructions. The 36 bp tags were aligned to the human genome using the ELAND algorithm. Figure shows the signal distribution along the complete length of chromosome 5 (figure A) and a zoomin to a 600 kb region (figure B). Figure C and D show the enrichment in two genomic regions on chromosome 3 and 12, respectively, containing EIF4A2 and GAPDH positive controls. The position of the amplicon used for validating the QPCR results is shown with an arrow.

Catalog #1: bs-53098R

Applications: ELISA, IF, dot-blot, ChIP-seq

Reactivity: Human, Mouse

Conjugate: Unconjugated

Western blot was performed on whole cell (25\u03bcg, lane 1) and histone extracts (15\u03bcg, lane 2) from HeLa cells, and on 1 \u03bcg of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using H4K5ac Polyclonal Antibody (bs-53099R). Antibody was diluted 1:500 in TBS-Tween containing 5% skim milk.
Dot Blot analysis was performed with peptides containing a variety of histone modifications and the unmodified H4. 0.2-100 pmol of the respective peptides were spotted on a membrane. H4K5ac Polyclonal Antibody (bs-53099R) was used at 1:5,000 dilution. Result shows high specificity of the antibody for the modification of interest.
ChIP were performed using HeLa cells using H4K5ac Polyclonal Antibody (bs-53099R). Chromatin was sheared from 1 million cells. A titration of the antibody consisting of 1, 2, 5, and 10\u03bcg per ChIP experiment was analyzed. IgG (2\u03bcg\/IP) was used as negative IP control. Quantitative PCR was performed with primers for promoter of the active gene EIF4A2 and for a region 1 kb upstream of the GAPDH as positive controls, and for the inactive MYOD1 gene and Sat2 satellite repeat region used as negative controls. Result shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).
ELISA was performed using a serial dilution of the\r\nH4K5ac Polyclonal Antibody (bs-53099R) against peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:8,900.

Catalog #1: bs-53099R

Applications: WB, ELISA, IHC-P, IF(ICC), dot-blot

Reactivity: Human, Mouse

Conjugate: Unconjugated

HeLa cells were stained with the Bioss antibody against H4K20me1 (Cat. bs-53104R) and with DAPI. Cells were fixed with 4% formaldehyde for 10\u2019 and blocked with PBS\/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labeled with the H4K20me1 antibody (left) diluted 1:500 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.
Western blot was performed on whole cell extracts (25 \u03bcg, lane 1) from HeLa cells, and on 1 \u03bcg of recombinant histone H2A, H2B, H3 and H4 (lane 2, 3, 4 and 5, respectively) using the Bioss antibody against H4K20me1 (cat. No. bs-53104R). The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The marker (in kDa) is shown on the left, the position of the protein is indicated on the right.
To check the specificity of the Bioss antibody against H4K20me1 (cat. No bs-53104R) a Dot Blot was performed with peptides containing different modifications of histone H3 and H4 or the unmodified H4K20 sequence. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. The figure shows a high specificity of the antibody for the modification of interest.
To determine the titer, an ELISA was performed using a serial dilution of the Bioss antibody directed against H4K20me1 (cat. No. bs-53104R) in antigen-coated wells. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:51,100.

Catalog #1: bs-53104R

Applications: WB, ELISA, IF(ICC), dot-blot

Reactivity: Human

Conjugate: Unconjugated

A Dot Blot analysis was performed to test the cross-reactivity of the Bioss antibody against H4K20me3 (Cat. No. bs-53115R) with peptides containing other histone modifications and the unmodified H4K20. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure shows a high specificity of the antibody for the modification of interest
Human osteosarcoma (U2OS) cells were stained with the Bioss antibody against H4K20me3 (Cat. No. bs-53115R) and with DAPI. Cells were fixed with ice-cold methanol for 10\u2019 and blocked with PBS\/TX-100 containing 5% normal goat serum. Figure A: cells were immunofluorescently labeled with the H4K20me3 antibody (left) diluted 1:300 in blocking solution followed by an anti-rabbit secondary antibody conjugated to Alexa568 or with DAPI (right). Figure B: staining of the cells with the H4K20me3 antibody after incubation of the antibody with blocking peptide at 5 ng\/\u03bcl.
To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Bioss antibody directed against H4K20me3 (Cat. No. bs-53115R), crude serum and flow through in antigen-coated wells. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:7,400.
ChIP assays were performed using HeLa cells, the Bioss antibody against H4K20me3 (Cat. No. bs-53115R) and optimized PCR primer sets for qPCR. ChIP was performed with an Auto Histone ChIP-seq kit with sheared chromatin from 1 million cells using the SX-8G IP-Star automated system. A titration of the antibody consisting of 1, 2, 5, and 10 \u03bcg per ChIP experiment was analyzed. IgG (1 \u03bcg\/IP) was used as negative IP control. QPCR was performed with primers for promoters of the active genes c-fos and GAPDH, used as negative controls, and for the Sat2 satellite repeat region used as a positive control. The figure shows the recovery expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

Catalog #1: bs-53115R

Applications: WB, ELISA, IHC-P, IF(ICC), IF, dot-blot, ChIP-seq

Reactivity: Human, Mouse, Others

Conjugate: Unconjugated

A Dot Blot analysis was performed to test the cross-reactivity of the Bioss antibody against H4K8ac (cat. No. bs-53133R) with peptides containing other histone modifications and the unmodified H4K8. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:20,000.
Histone extracts of HeLa cells (15 \u03bcg) were analyzed by Western blot using the Bioss antibody directed against H4K8ac (cat. No. bs-53133R) diluted 1:200 in TBS-Tween containing 5% skimmed milk.
To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Bioss antibody directed against H4K8ac (cat. No. bs-53133R), crude serum and flow through in antigen-coated wells. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution, the titer of the purified antibody was estimated to be 1:16,700.
Mouse NIH3T3 cells were stained with H4K8ac Polyclonal Antibody (cat. No. bs-53133R) and with DAPI. Cells were fixed with 4% formaldehyde for 10\u2019 and blocked with PBS\/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labeled with the H4K8ac antibody (left) diluted 1:500 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.

Catalog #1: bs-53133R

Applications: WB, ELISA, IF, dot-blot

Reactivity: Human, Mouse

Conjugate: Unconjugated

HeLa cells were stained with the Bioss antibody against H4K5,8,12ac (cat. No. bs-53096R) and with DAPI. Cells were fixed with 4% formaldehyde for 10\u2019 and blocked with PBS\/TX-100 containing 5% normal goat serum and 1% BSA. Figure A: cells were immunofluorescently labeled with the H4K5,8,12ac antibody (left) diluted 1:500 in blocking solution followed by Alexa488 conjugated secondary antibody. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right. Figure B, C, D, and E: staining of the cells with the H4K5,8,12ac antibody after incubation of the antibody with 10 ng\/\u03bcl of the following blocking peptides: H4K5,8,12 unmodified (figure B), H4K5,8,12ac (figure C), H2A.ZK5,7,11ac (figure D) and H4K5,8,12,16ac (figure E).
Western blot was performed on whole cell (25 \u03bcg, lane 1) and histone extracts (15 \u03bcg, lane 2) of HeLa cells, and on 1 \u03bcg of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the Bioss antibody against H4K5,8,12ac (cat. No. bs-53096R). The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk.
To test the cross-reactivity of the Bioss antibody against H4K5,8,12ac (cat. No. bs-53096R), a Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H4. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure shows a high specificity of the antibody for the modification of interest.
To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Bioss antibody directed against H4K5,8,12ac (cat. No. bs-53096R) in antigen-coated wells. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:14,500.

Catalog #1: bs-53096R

Applications: WB, ELISA, IP, IF, dot-blot, ChIP-seq

Reactivity: Human, Mouse

Conjugate: Unconjugated

Histone extracts (15 \u03bcg) from HeLa cells were analysed by Western blot using H3K4un (3C5) Monoclonal Antibody (Cat. No. bsm-53012M) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.
HeLa cells were stained with H3K4un (3C5) Monoclonal Antibody (Cat. No. bsm-53012M) and with DAPI. Cells were fixed with 4% formaldehyde for 10\u2019 and blocked with PBS\/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labelled with the H3K4un antibody (left) diluted 1:500 in blocking solution followed by an anti-mouse antibody conjugated to Alexa594. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.
To test the specificity an ELISA was performed using a serial dilution of H3K4un (3C5) Monoclonal Antibody (Cat. No. bsm-53012M). The wells were coated with peptides containing the unmodified H3K4 region as well as the mono-, di- and trimethylated H3K4. Figure 2 shows a high specificity of the antibody for the unmodified peptide.
ChIP assays were performed using HeLa cells, the monoclonal antibody against H3K4un (Cat. No. bsm-53012M) and optimized PCR primer sets for qPCR. A titration of the antibody consisting of 1, 5 and 10 \u03bcg per ChIP experiment was analysed. IgG (5 \u03bcg\/IP) was used as negative IP control. QPCR was performed with primers for the GAPDH promoter and for exon 2 of the myoglobin gene. Figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

Catalog #1: bsm-53012M

Applications: WB, ELISA, IF(ICC)

Reactivity: Human

Conjugate: Unconjugated

Histone extracts (15 \u03bcg) from HeLa cells were analysed by Western blot using the H3K4me1 Monoclonal Antibody (bsm-53013M) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the molecular weight marker (in kDa) is shown on the left.
HeLa cells were stained with H3K4me1 (34A1) Monoclonal Antibody (Cat. No. bsm-53013M) and with DAPI. Cells were fixed with 4% formaldehyde for 10\u2019 and blocked with PBS\/TX-100 containing 5% normal goat serum and 1% BSA. The cells were labelled with the H3K4me1 antibody (left) diluted 1:500 in blocking solution followed by an anti-mouse antibody conjugated to Alexa594. The middle panel shows staining of the nuclei with DAPI. A merge is shown on the right.
To test the specificity an ELISA was performed using a serial dilution of H3K4me1 (34A1) Monoclonal Antibody (Cat. No. bsm-53013M). The wells were coated with peptides containing the unmodified H3K4 as well as the mono-, di- and trimethylated H3K4 and the monomethylated H3K9. Figure shows a high specificity of the antibody for the modification of interest.
ChIP assays were performed using HeLa cells, H3K4me1 (34A1) Monoclonal Antibody (bsm-53013M) and optimized PCR primer sets for qPCR. ChIP was performed using sheared chromatin from 1.6 million cells. A titration of the antibody consisting of 1, 5 and 10 \u03bcg per ChIP experiment was analyzed. IgG (5 \u03bcg\/IP) was used as negative IP control. QPCR was performed with primers for the promoter and the coding region of the GAPDH gene, and for the RPL10 and HBB promoters. Figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

Catalog #1: bsm-53013M

Applications: WB, ELISA, IHC-P, IF(ICC)

Reactivity: Human

Conjugate: Unconjugated

Histone extracts (15 \u03bcg) from HeLa cells were analysed by Western blot using H3K4me2 (4B4) Monoclonal Antibody (cat. No. bsm-53014M) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.
HeLa cells were stained with H3K4me2 (4B4) Monoclonal Antibody (cat. No. bsm-53014M) and with DAPI. Cells were fixed with 4% formaldehyde for 10\u2019 and blocked with PBS\/TX-100 containing 5% normal goat serum and 1% BSA. The cells were labelled with the H3K4me2 antibody (left) diluted 1:500 in blocking solution followed by an anti-mouse antibody conjugated to Alexa594. The middle panel shows staining of the nuclei with DAPI. A merge of the two staining is shown on the right.
To test the specificity an ELISA was performed using a serial dilution of the H3K4me2 (4B4) Monoclonal Antibody (cat. No. bsm-53014M). The wells were coated with peptides containing the unmodified H3K4 as well as the mono-, di- and trimethylated H3K4 and the dimethylated H3K9. Figure shows a high specificity of the antibody for the modification of interest.
ChIP assays were performed using HeLa cells, H3K4me2 (4B4) Monoclonal Antibody (cat. No. bsm-53014M) and optimized PCR primer sets for qPCR. ChIP was performed using sheared chromatin from 1.6 million cells. A titration of the antibody consisting of 1, 5 and 10 \u03bcg per ChIP was analysed. IgG (5 \u03bcg\/IP) was used as negative IP control. QPCR was performed with primers for the promoter and the coding region of the GAPDH gene, and for the RPL10 and HBB promoters. Figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

Catalog #1: bsm-53014M

Applications: WB, ELISA, IF(ICC)

Reactivity: Human, Others

Conjugate: Unconjugated

Histone extracts (15 \u03bcg) from HeLa cells were analysed by Western blot using H3K4me3 (2G10) Monoclonal Antibody (Cat. No. bsm-53015M) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.
HeLa cells were stained with H3K4me3 (2G10) Monoclonal Antibody (Cat. No. bsm-53015M) and with DAPI. Cells were fixed with 4% formaldehyde for 10\u2019 and blocked with PBS\/TX-100 containing 5% normal goat serum and 1% BSA. The cells were labelled with the H3K4me3 antibody (left) diluted 1:500 in blocking solution followed by an anti-mouse antibody conjugated to Alexa594. The middle panel shows staining of the nuclei with DAPI. A merge of the two staining is shown on the right.
To test the specificity an ELISA was performed using a serial dilution of H3K4me3 (2G10) Monoclonal Antibody (Cat. No. bsm-53015M). The wells were coated with peptides containing the unmodified H3K4 as well as the mono-, di- and trimethylated H3K4 and the trimethylated H3K9. Figure shows a high specificity of the antibody for the modification of interest.
ChIP assays were performed using human HeLa cells, \r\nH3K4me3 (2G10) Monoclonal Antibody (bsm-53015M) and optimized PCR primer pairs for qPCR. ChIP was performed using sheared chromatin from 1 million cells on the SX-8G IP-Star automated system. A titration consisting of 1, 2, 5 and 10 \u03bcg of antibody per ChIP experiment was analyzed. IgG (2 \u03bcg\/IP) was used as a negative IP control. Quantitative PCR was performed with primers specific for the promoter of GAPDH and c-fos genes (positive controls), and exon 2 of the inactive myoglobin (MB) gene and Sat2 satellite repeat (negative controls). Figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). These results are in accordance with the observation that trimethylation of K4 at histone H3 is associated with the promoters of active genes.

Catalog #1: bsm-53015M

Applications: WB, ELISA, IF(ICC)

Reactivity: Human, Others

Conjugate: Unconjugated

To test the specificity an ELISA was performed using a serial dilution using 3K36me2 (6C9) Monoclonal Antibody (bsm-53020M). The wells were coated with peptides containing the unmodified H3K36 region as well as the mono-, di- and trimethylated H3K36 and the trimethylated H3K9. Figure shows a high specificity of the antibody for the peptide containing the modification of interest.
ChIP assays were performed using HeLa cells, H3K36me2 (6C9) Monoclonal Antibody (bsm-53020M) and optimized PCR primer pairs for qPCR. ChIP was performed using sheared chromatin from 1 million cells. A titration consisting of 0.5, 1, 2 and 5 \u03bcg of antibody per ChIP experiment was analyzed. IgG (1 \u03bcg\/IP) was used as a negative IP control. Quantitative PCR was performed with primers specific for a genomic region upstream of the TGM2 gene, used as a positive control, and for the inactive MYOD1 gene and the Sat2 satellite repeat, used as negative controls. Figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

Catalog #1: bsm-53020M

Applications: ELISA, IF(ICC)

Reactivity: Human

Conjugate: Unconjugated

Histone extracts (15 \u03bcg) from HeLa cells were analysed by Western blot using H3K9me2 (9B3) Monoclonal Antibody (Cat. No. bsm-53016M) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.
HeLa cells were stained with H3K9me2 (9B3) Monoclonal Antibody (bsm-53016M). Cells were fixed with 4% formaldehyde for 10\u2019 and blocked with PBS\/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labelled with the H3K9me2 antibody (left) diluted 1:500 in blocking solution followed by an anti-mouse antibody conjugated to Alexa594. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.
To test the specificity an ELISA was performed using a serial dilution H3K9me2 (9B3) Monoclonal Antibody (bsm-53016M). The wells were coated with peptides containing the unmodified H3K9 as well as the mono-, di- and trimethylated H3K9 and the dimethylated H3K4 and H3K27. Figure shows a high specificity of the antibody for the modification of interest.
ChIP assays were performed using HeLa cells, H3K9me2 (9B3) Monoclonal Antibody (bsm-53016M) and optimized PCR primer sets for qPCR. ChIP was performed using sheared chromatin from 1.6 million cells. A titration of the antibody consisting of 1, 3 and 9 \u03bcg per ChIP experiment was analysed. IgG (5 \u03bcg\/IP) was used as negative IP control. QPCR was performed with primers for the promoter and the coding region of the GAPDH gene, and for the RPL10 and HBB promoters. Figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

Catalog #1: bsm-53016M

Applications: WB, ELISA, IF(ICC), IF

Reactivity: Human

Conjugate: Unconjugated

Histone extracts (15 \u03bcg) from HeLa cells were analysed by Western blot using H3K27me3 (9B7) Monoclonal Antibody (bsm-53019M) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.
HeLa cells were stained with H3K27me3 (9B7) Monoclonal Antibody (bsm-53019M) and DAPI. Cells were fixed with 4% formaldehyde for 10\u2019 and blocked with PBS\/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labelled with the H3K27me3 antibody (left) diluted 1:500 in blocking solution followed by an anti-mouse secondary antibody conjugated to Alexa594. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.
To test the specificity an ELISA was performed using a serial dilution of H3K27me3 (9B7) Monoclonal Antibody (bsm-53019M). The wells were coated with peptides containing the unmodified H3K27 region as well as the mono-, di- and trimethylated H3K27 and the trimethylated H3K9. Figure shows a high specificity of the antibody for the peptide containing the modification of interest.
ChIP assays were performed using HeLa cells, H3K27me3 (9B7) Monoclonal Antibody (bsm-53019M) and optimized PCR primer sets for qPCR. ChIP was performed on sheared chromatin from 1 million cells. A titration of the antibody consisting of 1, 2, 5, and 10 \u03bcg per ChIP experiment was analysed. IgG (2 \u03bcg\/IP) was used as negative IP control. QPCR was performed with primers for the promoters of the active genes c-fos and GAPDH as negative controls, and for the coding regions of the inactive genes MYT1 and TSH2B as positive controls. Figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). These results are in accordance with the observation that H3K27me3 is preferably present at inactive genes.

Catalog #1: bsm-53019M

Applications: WB, ELISA, IF(ICC), IF

Reactivity: Human

Conjugate: Unconjugated

HeLa cells were stained with H3K27ac (1A3) Monoclonal Antibody (bsm-53022M) and with DAPI. Cells were fixed with 4% formaldehyde for 10\u2019 and blocked with PBS\/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labelled with the H3K27ac antibody (left) diluted 1:500 in blocking solution followed by an anti-mouse antibody conjugated to Alexa594. The middle panel shows staining of the nuclei with DAPI. A merge of the two staining is shown on the right.
To test the specificity an ELISA was performed using a serial dilution of H3K27ac (1A3) Monoclonal Antibody (bsm-53022M). The wells were coated with peptides containing the unmodified H3K27 region as well as the acetylated H3K27 and the acetylated H3K9. Figure shows a high specificity of the antibody for the peptide containing the modification of interest.
ChIP assays were performed using HeLa cells, H3K27ac (1A3) Monoclonal Antibody (bsm-53022M) and optimized PCR primer pairs for qPCR. ChIP was performed using sheared chromatin from 1 million cells. A titration consisting of 0.5, 1, 2 and 5 \u00b5g of antibody per ChIP experiment was analyzed. IgG (1\u00b5g\/IP) was used as a negative IP control. Quantitative PCR was performed with primers specific for the promoters of the EIF4A2 and GAPDH genes, used as positive controls, and for the inactive MYOD1 gene and the Sat2 satellite repeat, used as negative controls. Figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

Catalog #1: bsm-53022M

Applications: WB, ELISA, IF(ICC)

Reactivity: Human

Conjugate: Unconjugated

HeLa cells were stained with H3K36me3 (11D1) Monoclonal Antibody (bsm-53021M) and DAPI. Cells were fixed with methanol and blocked with PBS\/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labelled with the H3K36me3 antibody (left) diluted 1:500 in blocking solution followed by an anti-mouse antibody conjugated to Alexa594. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.
To test the specificity an ELISA was performed using a serial dilution of H3K36me3 (11D1) Monoclonal Antibody (bsm-53021M). The wells were coated with peptides containing unmodified H3K36 region as well as the mono-, di- and trimethylated H3K36 and the trimethylated H3K9. Figure shows a high specificity of the antibody for the peptide containing the modification of interest.
ChIP assays were performed using HeLa cells, H3K36me3 (11D1) Monoclonal Antibody (bsm-53021M) and optimized PCR primer pairs for qPCR. ChIP was performed using sheared chromatin from 1 million cells. A titration consisting of 0.5, 1, 2 and 5 \u03bcg of antibody per ChIP experiment was analyzed. IgG (1 \u03bcg\/IP) was used as a negative IP control. Quantitative PCR was performed with primers specific for the coding regions of the active EIF2S3 and CCT5 genes, used as positive controls, and for the inactive MYOD1 gene and the Sat2 satellite repeat, used as negative controls. Figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

Catalog #1: bsm-53021M

Applications: WB, ELISA, IF

Reactivity: Human

Conjugate: Unconjugated

Western blot was performed on histone extracts from HeLa cells (15 \u03bcg) with the H3K79me1 antibody (bs-53049R), diluted 1:1,000 and 1:2000 in TBS-Tween containing 5% skimmed milk. The molecular weight marker (Bio-Rad, broad range biotinylated SDS-PAGE standard) is shown on the left, the location of the protein of interest is indicated on the right.
A Dot Blot analysis was performed to test the cross reactivity of the H3K79me1 antibody (bs-53049R) with peptides containing other modifications of histone H3. These include di- and trimethylation of the same lysine and mono-, di- and trimethylation of lysine 9, 27 and 36. One hundred to 0.2 pmol of the peptides were spotted on a membrane. The antibody was used at a dilution of 1:100,000. The figure shows a high specificity of the antibody for the modification of interest.
To determine the titer of the antibody, an ELISA was performed using a serial dilution of the H3K79me1 antibody (bs-53049R) in antigen coated wells. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:30,000.
ChIP assays were performed using HeLa cells, H3K79me1 antibody (bs-53049R) and optimized PCR primer pairs for qPCR. Chromatin from 1.6 million cells were sheared. ChIP was performed with IgG (5 \u03bcg\/IP) as a negative IP control. The IP\u2019d DNA was analysed by qPCR using primers for different positive and negative loci. The results are expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). The figure shows recovery of the GAPDH promoter and myoglobin exon 2 with a titration of the H3K79me1 antibody consisting of 5, 10 and 15 \u03bcl per ChIP experiment.

Catalog #1: bs-53049R

Applications: WB, ELISA, dot-blot

Reactivity: Human

Conjugate: Unconjugated

Histone extracts of HeLa cells (15 \u03bcg) were analyzed by Western blot using the antibody against H3K4me2 (bs-53036R) diluted 1:750 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.
A Dot Blot analysis was performed to test the cross-reactivity of the antibody against H3K4me2 (bs-53036R) with peptides containing other modifications or unmodified sequences of histone H3. Other histone modifications include mono- and trimethylation of the same lysine and mono-, di- and trimethylation of lysine 9, 27 and 36 and 79. One hundred to 0.2 pmol of the peptides were spotted on a membrane. The antibody was used at a dilution of 1:20,000. The figure shows a high specificity of the antibody for the modification of interest.
To determine the titer, an ELISA was performed using a serial dilution of the antibody directed against H3K4me2 (bs-53036R). The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:2,600.
ChIP assays were performed using U2OS cells, the antibody against H3K4me2 (bs-53036R) and optimized PCR primer sets for qPCR. ChIP was performed using sheared chromatin from 1.6 million cells. A titration of the antibody consisting of 2, 5, 10 or 15 \u03bcl per ChIP experiment was analyzed. IgG (5 \u03bcg\/IP) was used as negative IP control. Quantitative PCR was performed with primers for the promoter of the ALDOA gene and for the coding region of the myogenic differentiation gene (MYOD), a gene that is inactive under normal conditions. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). These results are in accordance with the observation that dimethylation of K4 at histone H3 is more present in active genes than in silent genes.

Catalog #1: bs-53036R

Applications: WB, ELISA, IF, dot-blot

Reactivity: Human, Mouse

Conjugate: Unconjugated

ChIP assays were performed using U2OS cells, the antibody against H3K4me1 (bs-53037R) and optimized PCR primer sets for qPCR. ChIP was performed using sheared chromatin from 1.6 million cells. A titration of the antibody consisting of 2, 5, 10 or 15 \u03bcl per ChIP experiment was analyzed. IgG (5 \u03bcg\/IP) was used as negative IP control. Quantitative PCR was performed with primers for the promoter of the ALDOA gene and for the coding region of the myogenic differentiation gene (MYOD), a gene that is inactive under normal conditions. The figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).
To determine the titer, an ELISA was performed using a serial dilution of the antibody directed against H3K4me1 (bs-53037R). The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:3,800.
Histone extracts of HeLa cells (15 \u03bcg) were analyzed by Western blot using H3K4me1 Polyclonal Antibody (bs-53037R) diluted 1:750 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.
A Dot Blot analysis was performed to test the cross-reactivity of the antibody against H3K4me1 (bs-53037R) with peptides containing other modifications or unmodified sequences of histone H3. Other histone modifications include di- and trimethylation of the same lysine and mono-, di- and trimethylation of lysine 9, 27 and 36 and 79. One hundred to 0.2 pmol of the peptides were spotted on a membrane. The antibody was used at a dilution of 1:20,000. The figure shows a high specificity of the antibody for the modification of interest.

Catalog #1: bs-53037R

Applications: WB, ELISA, dot-blot, ChIP-seq

Reactivity: Human

Conjugate: Unconjugated

Histone extracts of HeLa cells (15 \u03bcg) were analysed by Western blot using the H3K79me2 antibody (bs-53039R) diluted 1:250 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the left; the marker (in kDa) is shown on the right.
A Dot Blot analysis was performed to test the cross reactivity of the H3K79me2 antibody (bs-53039R) with peptides containing other modifications of histone H3. Other H3 modifications include mono- and trimethylation of the same lysine and mono-, di- and trimethylation of lysine 9, 27 and 36. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:50,000. The figure shows a high specificity of the antibody for the modification of interest.
ChIP assays were performed using human osteosarcoma (U2OS) cells, the H3K79me2 antibody (bs-53039R) and optimized PCR primer sets for qPCR. ChIP was performed using sheared chromatin from 1.6 million cells. A titration of the antibody consisting of 2, 7 and 15 \u03bcl per ChIP experiment was analysed. Additionally, ChIP was performed with 2 and 15 \u03bcl of antibody after incubation with 5 nmol blocking peptide for 1 hour at room temperature. IgG (5 \u03bcg\/IP) was used as negative IP control. QPCR was performed with primers for the ALDOA (fructose-bisphosphate aldolase A) promoter and for the coding region of the myogenic differentiation gene (MYOD), a gene that is inactive at normal conditions. The figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).
To determine the titer, an ELISA was performed using a serial dilution of the H3K79me2 antibody (bs-53039R). The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution the titer of the antibody was estimated to be 1:4,500.

Catalog #1: bs-53039R

Applications: WB, ELISA, dot-blot

Reactivity: Human

Conjugate: Unconjugated

ChIP was performed with 5 \u03bcl of the antibody against H3K36me3 (bs-53041R) on sheared chromatin from 1 million HeLaS3 cells. IgG (2 \u03bcg\/IP) was used as a negative IP control. The IP\u2019d DNA was analysed by QPCR with optimized PCR primer pairs for the coding and promoter region of the active GAPDH gene, for the coding region of the inactive TSH2B gene and for the Sat2 satellite repeat. The IP\u2019d DNA was subsequently analysed with an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer\u2019s instructions. The 36 bp tags were aligned to the human genome using the ELAND algorithm. The figure shows the results in 200 kb regions of chromosome 12 (including the GAPDH positive control), 6 and 7 and 14. These results clearly show an enrichment of the H3K36me3 at active genes
ChIP assays were performed using human osteosarcoma (U2OS) cells, the antibody against H3K36me3 (bs-53041R) and optimized PCR primer sets for qPCR. ChIP was performed using sheared chromatin from 1.6 million cells. A titration of the antibody consisting of 2, 5, 10 and 15 \u03bcl per ChIP experiment was analyzed. IgG (5 \u03bcg\/IP) was used as a negative IP control. Quantitative PCR was performed using primer sets for the housekeeping gene GAPDH and for myogenic differentiation gene (MYOD). The figure shows the recovery expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis.
To determine the titer, an ELISA was performed using a serial dilution of the antibody directed against H3K36me3 (bs-53041R). The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:12,700.
A Dot Blot analysis was performed to test the cross-reactivity of the antibody against H3K36me3 (bs-53041R) with peptides containing other modifications of histone H3. Other histone modifications include mono- and dimethylation of the same lysine and mono-, di- and trimethylation of lysine 9, 27 and 79. One hundred to 0.2 pmol of peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:100,000. The figure shows a high specificity of the antibody for the modification of interest.

Catalog #1: bs-53041R

Applications: WB, ELISA, dot-blot, ChIP-seq

Reactivity: Human, Mouse

Conjugate: Unconjugated

HeLa cells were stained with the H3K9me1 antibody (bs-53043R) and with DAPI. Cells were formaldehyde fixated, permeabilized with Triton X-100 and blocked with PBS containing 2.5% BSA. Figure A: cells were immunofluorescently labelled with the H3K9me1 antibody (diluted 1:200 and incubated for 1 hour at room temperature) followed by goat anti-rabbit antibody conjugated to DyLight. Figure B: staining of the nuclei with DAPI, which specifically labels DNA. Both antibody and DAPI staining are restricted to the nucleus.
To determine the titer, an ELISA was performed using a serial dilution of the H3K9me1 antibody (bs-53043R). The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution the titer of the antibody was estimated to be 1:50,000.
A Dot Blot analysis was performed to test the cross reactivity of H3K9me1 antibody (bs-53043R) with peptides containing other modifications of histone H3. Other histone modifications include di- and trimethylation of the same lysine and mono-, di- and trimethylation of lysine 27, 36 and 79. One hundred to 0.2 pmol of peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:200,000. The figure shows a high specificity of the antibody for the modification of interest.
Histone extracts of HeLa cells (15 \u03bcg) were analysed by Western blot using the H3K9me1 antibody (bs-53043R) diluted 1:1000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the left; the marker (in kDa) is shown on the right.

Catalog #1: bs-53043R

Applications: WB, ELISA, IF, dot-blot

Reactivity: Human

Conjugate: Unconjugated

HeLa cells were treated with TSA (figure on left) or with colcemid (figure on right), and 15 \u03bcg of histone extracts of these cells were analyzed by Western blot using the antibody against H3S10p (bs-53069R) diluted 1:500 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the left; the marker (in kDa) is shown on the right. The result of the Western analysis with the antibody is shown in lane 1; lane 2 shows the same analysis after incubation of the antibody with 750 pmol blocking peptide for 1 hour at room temperature.
Hela asynchronous cells were stained with the antibody against H3S10p (bs-53069R) and with DAPI. Cells were fixed with formaldehyde, permeabilized with sodium citrate and Triton X100 and blocked with PBS containing 2.5% BSA. (left) Cells were immunofluorescently labeled with the H3S10p antibody (diluted 1:200 and incubated for 1 hour at room temperature) followed by goat anti-rabbit antibody conjugated to DyLight 488. (right) The nuclei were stained with DAPI, which specifically labels DNA. Phosphorylation of H3 on serine 10 occurs on condensed chromosomes during mitosis. This explains the dense staining of one of the cells (indicated by an arrow) in the figure.
A Dot Blot analysis was performed to test the cross-reactivity of the antibody against H3S10p (bs-53069R) with peptides containing other modifications of histone H3 and H4 and with peptides containing unmodified sequences from histone H3. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. The figure shows a high specificity of the antibody for the modification of interest. Note that the antibody does not recognize the H3S10p modification if the H3K9ac modification is present.
To determine the titer, an ELISA was performed using a serial dilution of the antibody directed against human H3S10p (bs-53069R). The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:35,000

Catalog #1: bs-53069R

Applications: WB, ELISA, IF, dot-blot

Reactivity: Human

Conjugate: Unconjugated

ChIP assays were performed using HeLa cells treated with colcemid, the H3K27me3S28p antibody (bs-53053R) and optimized PCR primer pairs for qPCR. ChIP was performed using sheared chromatin from 10,000 cells. A titration of the antibody consisting of 1, 5 and 10 \u03bcl per ChIP experiment was analysed. Additionally, ChIP was performed after incubation of the antibody with 5 nmol blocking peptide (cat. No. sp-091-050) for 1 hour at room temperature. IgG (5 \u03bcg\/IP) was used as negative IP control. QPCR was performed with primers for the promoters of the active genes c-fos and RPL30 and for the inactive gene MYOD. The figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).
To determine the titer, an ELISA was performed using a serial dilution of the H3K27me3S28p antibody (bs-53053R). The antigen used was a peptide containing the histone modifications of interest. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:8,300.
A Dot Blot analysis was performed to test the cross reactivity of the H3K27me3S28p antibody (bs-53053R) with peptides containing other modifications of histone H3 and H4 and unmodified sequences from histone H3. One hundred to 0.2 pmol of the peptides were spotted on a membrane. The antibody was used at a dilution of 1:20,000. The figure shows a high specificity of the antibody for the peptide containing the modifications of interest.
Hela cells were stained with the H3K27me3S28p antibody (bs-53053R) and with DAPI. Cells were fixed with formaldehyde, permeabilized with sodium citrate and Triton X100 and blocked with PBS containing 2.5% BSA. Left side: cells were immunofluorescently labelled with the H3K27me3S28p antibody (diluted 1:200 and incubated for 1 hour at room temperature) followed by goat anti-rabbit antibody conjugated to DyLight 488. Right side: the nuclei were stained with DAPI, which specifically labels DNA. Phosphorylation of H3 on serine 28 is increased during late G2 phase and reaches a maximum in metaphase cells. This may explain the different staining intensities of different cells.

Catalog #1: bs-53053R

Applications: WB, ELISA, IP, IF, dot-blot, ChIP-seq

Reactivity: Human

Conjugate: Unconjugated

ChIP assays were performed using human osteosarcoma (U2OS) cells, the antibody against H3R17me2 (bs-53054R) and optimized PCR primer sets for qPCR. ChIP was performed using sheared chromatin from 1.6 million cells per ChIP reaction. A titration of the antibody consisting of 2, 5, 10 and 15 \u03bcl per ChIP experiment was analyzed. IgG (5 \u03bcg\/IP) was used as negative IP control. The figures show the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). Left Figure: QPCR performed with primers for the GAPDH promoter and for exon 2 of the myoglobin gene. Right Figure: QPCR performed with primers for the promoter of the active ALDOA gene and for the coding region of the inactive MYOD gene.
To determine the titer, an ELISA was performed using a serial dilution of the antibody directed against H3R17me2 (bs-53054R). The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:40,000.
A Dot Blot analysis was performed to test the cross-reactivity of the antibody against H3R17me2 (bs-53054R) with peptides containing other modifications of histone H3 and H4 and unmodified sequences from histone H3. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. The figure shows a high specificity of the antibody for the modification of interest.
Histone extracts of HeLa cells (15 \u03bcg) were analyzed by Western blot using the antibody against H3R17me2 (bs-53054R) diluted 1:250 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the left; the marker (in kDa) is shown on the right.

Catalog #1: bs-53054R

Applications: WB, ELISA, dot-blot

Reactivity: Human

Conjugate: Unconjugated

Histone extracts of HeLa cells (15 \u03bcg) were analysed by Western blot using the H3K9acS10p antibody (bs-53062R) diluted 1:250 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the left; the marker (in kDa) is shown on the right.
A Dot Blot analysis was performed to test the cross reactivity of the H3K9acS10p antibody (bs-53062) with peptides containing other modifications of histone H4 and H3 or unmodified histone H3 sequences. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. The figure shows a high specificity of the antibody for the double modification.
Immunofluorescence using the H3K9acS10p antibody at 1:500 dilution in Hela cells.
ChIP assays were performed using human osteosarcoma (U2OS) cells, the H3K9acS10p antibody (bs-53062R) and optimized PCR primer sets for qPCR. ChIP was performed using sheared chromatin from 1.5 million cells. A titration of the antibody consisting of 5 and 15 \u03bcl per ChIP experiment was analysed. IgG (5 \u03bcg\/IP) was used as negative IP control. QPCR was performed with primers for the ALDOA (fructose-bisphosphate aldolase A) promoter and for the coding region of the myogenic differentiation gene (MYOD), a gene that is inactive at normal conditions. The figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

Catalog #1: bs-53062R

Applications: WB, ELISA, IF, dot-blot

Reactivity: Human

Conjugate: Unconjugated

ChIP assays were performed using HeLa cells, the Bioss antibody against H3K9\/14ac (Cat. No. bs-53093R) and optimized primer pairs for qPCR. ChIP was performed with a HighCell# ChIP kit, using sheared chromatin from 1.5 million cells. A titration of the antibody consisting of 1, 2, 5 and 10 \u03bcg per ChIP experiment was analyzed. IgG (5 \u03bcg\/IP) was used as negative IP control.QPCR was performed using primers specific for the promoter of the ACTB gene as a positive control target and for exon 2 of the MB gene as a negative control target. The figure shows the recovery (the relative amount of immunoprecipitated DNA compared to input DNA). These results confirm the observation that acetylation of H3K9\/14 is present at active promoters.
ChIP was performed with 1 \u03bcg of H3K9\/14ac Polyclonal Antibody (bs-53093R) on sheared chromatin from 1 million HeLaS3 cells using an Auto Histone ChIP-seq kit on the IP-Star automated system. IgG (2 \u03bcg\/IP) was used as a negative IP control. The IP\u2019d DNA was analyzed by qPCR with optimized PCR primer pairs for the promoters of the active GAPDH and c-fos genes, used as positive control targets, and the coding region of the inactive MB gene and the Sat2 satellite repeat, used as negative control targets (figure A). The IP\u2019d DNA was subsequently analyzed with an Illumina Genome Analyzer. Library preparation, cluster generation, and sequencing were performed according to the manufacturer\u2019s instructions. The 36 bp tags were aligned to the human genome using the ELAND algorithm. The figure shows the peak distribution along the complete sequence and an 800 kb region of the X-chromosome (figure B and C) and in 100 kb regions surrounding the RBM3, GAPDH and c-fos genes (figure D, E, and F). These results clearly show an enrichment of the H3K9\/14 double acetylation at the promoters of active genes.
To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Bioss antibody directed against H3K9\/14ac (Cat. No. bs-53093R), crude serum and flow through in antigen-coated wells. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution, the titer of the purified antibody was estimated to be 1:5,900.
A Dot Blot analysis was performed to test the cross-reactivity of the Bioss antibody against H3K9\/14ac (Cat. No. bs-53093R) with peptides containing other histone modifications and the unmodified H3K9\/14 sequence. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:20,000. The figure shows a high specificity of the antibody for the modification of interest.

Catalog #1: bs-53093R

Applications: WB, ELISA, IF, dot-blot, ChIP-seq

Reactivity: Human, Mouse, Others

Conjugate: Unconjugated

ChIP was performed with 0.5 \u03bcl of the H3K36me2 antibody (bs-53071R) on sheared chromatin from 1 million HeLa cells. The IP\u2019d DNA was subsequently analyzed on an Illumina Genome Analyzer. Library preparation, cluster generation and sequencing were performed according to the manufacturer\u2019s instructions. The 36 bp tags were aligned to the human genome using the ELAND algorithm. The figure shows the signal distribution along 3 genomic regions of chromosome 20, 12 and X, respectively.
ChIP assays were performed using human HeLa cells, the H3K36me2 antibody (bs-53071R) and optimized PCR primer sets for qPCR. ChIP was performed using sheared chromatin from 10,000 cells. A titration of the antibody consisting of 1, 5, and 10 \u03bcl per ChIP experiment was analyzed. Additionally, the same titration was analyzed after incubation of the antibody with 5 nmol blocking peptide for 1 hour at room temperature. IgG (5 \u03bcg\/IP) was used as negative IP control. QPCR was performed with primers for the promoter of the active genes GAPDH and ALDOA and for the coding region of the myogenic differentiation gene (MYOD). The figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).
To determine the titer, an ELISA was performed using a serial dilution of the H3K36me2 antibody (bs-53071R). The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:31,000.
Histone extracts of HeLa cells (15 \u03bcg) were analysed by Western blot using the H3K36me2 antibody (bs-53071R) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left. The result of the Western analysis with the antibody is shown in lane 1; lane 2 shows the same analysis after incubation of the antibody with 5 nmol blocking peptide for 1 hour at room temperature.

Catalog #1: bs-53071R

Applications: WB, ELISA, IF, dot-blot, ChIP-seq

Reactivity: Human

Conjugate: Unconjugated

ChIP assays were performed using HeLa cells treated with colcemid, the antibody against H3K9me3S10p (bs-53072R) and optimized PCR primer sets for qPCR. ChIP was performed using sheared chromatin from 10,000 cells per IP. A titration of the antibody consisting of 1, 5, and 10 \u03bcl per ChIP experiment was analyzed. Additionally, the same titration was analyzed after incubation of the antibody with 5 nmol blocking peptide (cat. No. sp-128-050) for 1 hour at room temperature. IgG (5 \u03bcg\/IP) was used as negative IP control. QPCR was performed with primers for the promoter of the active genes GAPDH and c-fos and for the heterochromatin marker Sat2. The figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).
To determine the titer, an ELISA was performed using a serial dilution of the antibody directed against human H3K9me3S10p (bs-53072R). The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:87,000.
A Dot Blot analysis was performed to test the cross-reactivity of the antibody against H3K9me3S10p (bs-53072R) with peptides containing other modifications and unmodified sequences of histone H3. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:1,000. The figure shows a high specificity of the antibody for the modification of interest.
HeLa cells were treated with colcemid to block the cell cycle in metaphase and 15 \u03bcg of histone extracts of these cells were analyzed by Western blot with the antibody against H3K9me3S10p (bs-53072R) diluted 1:500 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left. The result of the Western analysis with the antibody is shown in lane 2; the same analysis after incubation of the antibody with 5 nmol blocking peptide for 1 hour at room temperature is shown in lane 1.

Catalog #1: bs-53072R

Applications: WB, ELISA, IF, dot-blot

Reactivity: Human

Conjugate: Unconjugated

Whole cell extracts from HeLa cells (25 \u03bcg) were analysed by Western blot using the H3pan antibody (bs-53074R) diluted 1:500 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.
ChIP assays were performed using human HeLa cells, the Diagenode antibody against H3pan (bs-53074R) and optimized PCR primer sets for qPCR. ChIP was performed using sheared chromatin from 1 million cells. A titration of the antibody consisting of 1, 2, 5, and 10 \u03bcl per ChIP experiment was analysed. IgG (2 \u03bcg\/IP) was used as negative IP control. QPCR was performed with primers for the promoters of the active GAPDH and EIF4A2 genes, used as negative controls, and for the inactive MYOD1 and the Sat2 satellite repeat, used as positive controls. The figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).
To determine the titer of the antibody, an ELISA was performed using a serial dilution of the H3pan antibody (bs-53074R). The plates were coated with the peptides used for immunization. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be >1:1,000,000.

Catalog #1: bs-53074R

Applications: WB, ELISA

Reactivity: Human, Mouse, Rat

Conjugate: Unconjugated

To determine the titer, an ELISA was performed using a serial dilution of the H3K23ac antibody (bs-53076R). The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:2,750.
A Dot Blot analysis was performed to test the cross reactivity of the H3K23ac antibody (bs-53076R) with with peptides containing other modifications and unmodified sequences of histone H3. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:1,000. The figure shows a high specificity of the antibody for the modification of interest.

Catalog #1: bs-53076R

Applications: ELISA, dot-blot

Reactivity: Human

Conjugate: Unconjugated

ChIP assays were performed using HeLa cells, H3K9ac Polyclonal Antibody (Cat. No. bs-53092R) and optimized PCR primer pairs for qPCR. ChIP was performed with a ChIP-seq kit, using sheared chromatin from 1 million cells. A titration consisting of 1, 2, 5 and 10 \u03bcg of antibody per ChIP experiment was analyzed. IgG (2 \u03bcg\/IP) was used as a negative IP control. Quantitative PCR was performed with primers specific for the promoter of the active genes GAPDH and EIF4A2, used as positive controls, and for exon 2 of the inactive myoglobin (MB) gene and the Sat2 satellite repeat, used as negative controls. The figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). These results are in accordance with the observation that acetylation of K9 at histone H3 is associated with the promoters of active genes.
ChIP was performed with 1 \u03bcg of the Bioss antibody against H3K9ac (Cat. No. bs-53092R) and the IP\u2019d DNA was subsequently analyzed on an Illumina Genome Analyzer. Library preparation, cluster generation, and sequencing were performed according to the manufacturer\u2019s instructions. The 36 bp tags were aligned to the human genome using the ELAND algorithm. The figure shows the peak distribution along the complete sequence and an 800 kb region of the X-chromosome (figure A and B) and in 100 kb regions surrounding the GAPDH and EIF4A2 positive control genes ( figure C and D). These results clearly show an enrichment of H3K9ac at the promoters of active genes.
To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Bioss antibody directed against H3K9ac (Cat. No. bs-53092R) in antigen-coated wells. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:31,700.
To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Bioss antibody directed against H3K9ac (Cat. No. bs-53092R) in antigen-coated wells. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:31,700.

Catalog #1: bs-53092R

Applications: WB, ELISA, IF, dot-blot, ChIP-seq

Reactivity: Human, Mouse, Others

Conjugate: Unconjugated

To determine the titer, an ELISA was performed using a serial dilution of the H3K18ac antibody (bs-53075R). The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:8,800.
A Dot Blot analysis was performed to test the cross reactivity of the H3K18ac antibody (bs-53075R) with with peptides containing other modifications and unmodified sequences of histone H3. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:1,000. The figure shows a high specificity of the antibody for the modification of interest.

Catalog #1: bs-53075R

Applications: ELISA, dot-blot

Reactivity: Human

Conjugate: Unconjugated

ChIP assays were performed using HeLa cells, the Bioss antibody against H3K27me1 (Cat. No. bs-53108R) and optimized PCR primer sets for qPCR. ChIP was performed using sheared chromatin from 100,000 cells. A titration of the antibody consisting of 1, 2, 5 and 10 \u03bcg per ChIP experiment was analyzed. IgG (2 \u03bcg\/IP) was used as negative IP control. QPCR was performed with primers for the promoter and the coding region of the active gene GAPDH used as a negative and a positive control target, respectively. The figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).
To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Bioss antibody directed against H3K27me1 (Cat. No. bs-53108R). The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution, the titer of the purified antibody was estimated to be 1:32,900.
A Dot Blot analysis was performed to test the cross-reactivity of the Bioss antibody against H3K27me1 (Cat. No. bs-53108R) with peptides containing other modifications and unmodified sequences of histone H3 and H4. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. The figure shows a high specificity of the antibody for the modification of interest.
Histone extracts (15 \u03bcg) from HeLa cells were analyzed by Western blot using the Bioss antibody against H3K27me1 (Cat. No. bs-53108R) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.

Catalog #1: bs-53108R

Applications: WB, ELISA, IF, dot-blot

Reactivity: Human, Others

Conjugate: Unconjugated

Histone extracts of HeLa cells (15 \u03bcg) were analyzed by Western blot using the Bioss antibody against H3K4me2 (Cat. No. bs-53105R) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.\r\n
A Dot Blot analysis was performed to test the cross-reactivity of the Bioss antibody against H3K4me2 (Cat. No. bs-53105R) with peptides containing other modifications of histone H3 and H4 and the unmodified H3K4 sequence. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure shows a high specificity of the antibody for the modification of interest.
To determine the titer, an ELISA was performed using a serial dilution of H3K4me2 Polyclonal Antibody (bs-53105R), crude serum and flow through in antigen coated wells. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:12,600.
ChIP was performed with the Bioss antibody against H3K4me2 (Cat. No. bs-53105R) on sheared chromatin from 1 million HeLaS3 cells using an Auto Histone ChIP-seq kit. A titration of the antibody consisting of 1, 2, 5 and 10 \u03bcg per ChIP experiment was analyzed. IgG (2 \u03bcg\/IP) was used as negative IP control. Quantitative PCR was performed with primers for the promoter and coding region of the active GAPDH gene, the promoter of the active c-fos gene and for the coding region of the inactive TSH2B. The figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

Catalog #1: bs-53105R

Applications: WB, ELISA, dot-blot, ChIP-seq

Reactivity: Human

Conjugate: Unconjugated

ChIP was performed with the Bioss antibody against H3K4me1 (Cat. No. bs-53106R) on sheared chromatin from 1 million HeLaS3 cells using a ChIP-seq kit. A titration of the antibody consisting of 1, 2, 5 and 10 \u03bcg per ChIP experiment was analyzed. IgG (2 \u03bcg\/IP) was used as negative IP control. Quantitative PCR was performed with primers for a region surrounding the ACTB and GAS2L1 genes, used as positive controls, and for the promoters of the GAPDH and EIF4A2 genes, used as negative controls. The figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).
ChIP was performed as described above with 1 \u03bcg of the Bioss antibody against H3K4me1 (Cat. No. bs-53106R). The IP\u2019d DNA was subsequently analyzed on an Illumina Genome Analyzer. Library preparation, cluster generation, and sequencing were performed according to the manufacturer\u2019s instructions. The 36 bp tags were aligned to the human genome using the ELAND algorithm. Figure A and B show the enrichment in chromosomal regions surrounding the ACTB and GAS2L1 positive control genes. The position of the amplicon used in the qPCR validation is indicated by an arrow. Figure C and D show the H3K4me1 signal in two 1 Mb regions of chromosome 5, and X respectively.
To determine the titer, an ELISA was performed using a serial dilution of the Bioss antibody directed against H3K4me1 (Cat. No. bs-53106R) in antigen-coated wells. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:20,100.
A Dot Blot analysis was performed to test the cross-reactivity of the Bioss antibody against H3K4me1 (Cat. No. bs-53106R) with peptides containing other modifications or unmodified sequences of histone H3 and H4. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:10,000. The figure shows a high specificity of the antibody for the modification of interest.

Catalog #1: bs-53106R

Applications: WB, ELISA, IF, dot-blot, ChIP-seq

Reactivity: Human, Mouse

Conjugate: Unconjugated

ChIP assays were performed using HeLa cells, the Bioss antibody against H3K27me2 (cat. No. bs-53109R) and optimized PCR primer sets for qPCR. ChIP was performed with an Auto Histone ChIP-seq kit on the SX-8G IP-Star Compact automated system, using sheared chromatin from 1 million cells. A titration of the antibody consisting of 1, 2, 5, and 10 \u03bcg per ChIP experiment was analyzed. IgG (2 \u03bcg\/IP) was used as negative IP control. QPCR was performed with primers for the promoter of the active GAPDH and EIF4A2 genes, used as negative controls, and for the promoter of the inactive HBB and the coding region of the inactive MYOD1 genes, used as positive controls. The figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). These results are in accordance with the observation that H3K27me2 is preferably present at silent genes.
To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Bioss antibody directed against H3K27me2 (cat. No. bs-53109R). The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:480,000.
A Dot Blot analysis was performed to test the cross-reactivity of the Bioss antibody against H3K27me2 (cat. No. bs-53109R) with peptides containing other modifications of histone H3 and H4 and the unmodified sequence. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:50,000. The figure shows a high specificity of the antibody for the modification of interest.
Histone extracts (15 \u03bcg) from HeLa cells were analyzed by Western blot using the Bioss antibody against H3K27me2 (cat. No. bs-53109R) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.

Catalog #1: bs-53109R

Applications: WB, ELISA, IF, dot-blot

Reactivity: Human

Conjugate: Unconjugated

Histone extracts (15 \u03bcg) from HeLa cells were analyzed by Western blot using the Bioss antibody against H3K9me1 (Cat. No. bs-53120R) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.
A Dot Blot analysis was performed to test the cross-reactivity of the Bioss antibody against H3K9me1 (bs-53120R) with peptides containing other modifications and the unmodified sequence of histone H3. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. The figure shows specificity of the antibody for the modification of interest.
To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Bioss antibody directed against human H3K9me1 (Cat. No. bs-53120R), crude serum and flow through in antigen-coated wells. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:68,000.
ChIP assays were performed using human HeLa cells, the Bioss antibody directed against H3K9me1 (Cat. No. bs-53120R) and optimized PCR primer sets for qPCR. ChIP was performed with a Low Cell# ChIP kit, using sheared chromatin from 10,000 cells. 1 and 5 \u03bcg of the antibody and 5 \u03bcg of IgG (negative IP control) were used per ChIP experiment. qPCR was performed with primers for the GAPDH promoter and for the inactive gene MYOD. The figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). These results are in accordance with the observation that H3K9me1 is preferably present at silent genes.

Catalog #1: bs-53120R

Applications: WB, ELISA, dot-blot

Reactivity: Human, Others

Conjugate: Unconjugated