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Primary Antibodies

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ChIP assays were performed using HeLa cells, the Bioss antibody against H3K79me2 (Cat. No. bs-53111R) and optimized PCR primer pairs for qPCR. ChIP was performed with an Auto Histone ChIP-seq kit, 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. 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 with 1 \u03bcg of the Bioss antibody against H3K79me2 (Cat. No. bs-53111R) as described above. The IP\u2019d DNA was 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 peak distribution along the complete sequence and a 5 Mb region of chromosome 1 (figure A and B) and in two 300 kb regions surrounding the EIF2S3 and CCT5 positive control genes (figure C and D).
To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Bioss antibody directed against H3K79me2 (Cat. No. bs-53111R). 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 1:6,600.
A Dot Blot analysis was performed to test the cross-reactivity of the Bioss antibody against H3K79me2 (Cat. No. bs-53111R) 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 1:5,000. The figure shows a high specificity of the antibody for the modification of interest.

Catalog #1: bs-53111R

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

Reactivity: Human

Conjugate: Unconjugated

Western blot was performed on whole cell (25 \u00b5g, lane 1) and histone extracts (15 \u00b5g, lane 2) from HeLa cells, and on 1 \u00b5g of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the Bioss antibody against H3K9me3 (Cat. No. bs-53114R). The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk.
HeLa cells were stained with the Bioss antibody against H3K9me3 (bs-53114R) 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 H3K9me3 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 both stainings is shown on the right.
To determine the titer of the antibody, an ELISA was performed using a serial dilution of the antibody directed against human H3K9me3 (bs-53114R) 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.
A Dot Blot analysis was performed to test the cross-reactivity of the antibody against H3K9me3 (Cat. No. bs-53114R) 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.

Catalog #1: bs-53114R

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

Reactivity: Human, Mouse, Others, Zebrafish

Conjugate: Unconjugated

Histone extracts (15 \u03bcg) from HeLa cells were analyzed by Western blot using the Bioss antibody against H3K9me2 (Cat. No. bs-53118R) 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 H3K9me2 (Cat. No. bs-53118R) with peptides containing other modifications of histone H3 and the unmodified H3K9 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. Figure shows a high specificity of the antibody for the modification of interest.
Mouse NIH3T3 cells were stained with the Bioss antibody against H3K9me2 (Cat. No. bs-53118R) 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 H3K9me2 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 determine the titer of the antibody, an ELISA was performed using a serial dilution of the Bioss antibody against H3K9me2 (Cat. No. bs-53118R), crude serum and Flow through. 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 1:103,000.

Catalog #1: bs-53118R

Applications: WB, ELISA, IF, dot-blot

Reactivity: Human, Mouse, Others

Conjugate: Unconjugated

A Dot Blot analysis was performed to test the cross-reactivity of the Bioss antibody against H3K36me3 (bs-53116R) with peptides containing other H3 and H4 modifications 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 1:20,000. 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 H3K36me3 (Cat. No. bs-53116R) and the crude serum. 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 1:19,300.
ChIP was performed with 2 \u03bcg of the Bioss antibody against H3K36me3 (bs-53116R) on sheared chromatin from 1 million HeLaS3 cells using a ChIP-seq kit. 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 promoter and coding region of the active GAPDH, for a region located 1 kb upstream of the GAPDH promoter and for the coding region of the active ACTB gene (figure A). 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 B shows the obtained profiles in genomic regions of chromosome 12 (including the GAPDH positive control), 7 (including the ACTB positive control), 14 and 3, respectively. These results clearly show an enrichment of the H3K36me3 at active genes.
ChIP assays were performed using HeLa cells, the Bioss antibody against H3K36me3 (Cat. No. bs-53116R) and optimized PCR primer sets 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. QPCR was performed with primers for the promoter and coding region of the active GAPDH, for a region located 1 kb upstream of the GAPDH promoter and for the Sat2 satellite repeat. 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-53116R

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

Reactivity: Human, Others, Zebrafish, Drosophila

Conjugate: Unconjugated

A Dot Blot analysis was performed to test the cross-reactivity of the Bioss antibody against H3S10p (Cat. No. bs-53135R) with peptides containing other modifications of histone H3 or the unmodified H3S10 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. Note that the antibody does not recognize the H3S10p modification if the neighboring K9 is acetylated or trimethylated.
HeLa cells were treated with colcemid which blocks the cell cycle in metaphase and 15 \u03bcg of histone extracts of the cells were analyzed by Western blot using the Bioss antibody against H3S10p (Cat. No. bs-53135R) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk.
To determine the titer, an ELISA was performed using a serial dilution of the Bioss antibody directed against H3S10p (Cat. No. bs-53135R) and the crude serum. 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 1:5,200.
Figure A: ChIP assays were performed using HeLa cells treated with colcemid to block the cells in metaphase, H3S10p Polyclonal Antibody (bs-53135R) and optimized PCR primer sets for qPCR. ChIP was performed with LowCell ChIP kit on sheared chromatin from 10,000 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 (5 \u03bcg\/IP) was used as negative IP control. QPCR was performed with primers for the promoter of the active genes c-fos and RPL30. Figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). Figure B: ChIP was performed as described above using 2 \u03bcg of H3S10p antibody and sheared chromatin from 10,000 HeLa cells treated with colcemid (sample 1) or from 10,000 untreated cells (sample 2). QPCR was performed with primers for the promoter of the active genes c-fos and RPL30, and for the Sat2 satellite repeat region.

Catalog #1: bs-53135R

Applications: WB, ELISA, IF, dot-blot

Reactivity: Human

Conjugate: Unconjugated

Human osteosarcoma (U2OS) cells were stained with the Bioss antibody against H3K79me3 (Cat. No. bs-53121R) and with DAPI. Cells were fixed with 2.5% formaldehyde for 30\u2019 and blocked with PBS\/TX-100 containing 1% BSA. Figure A: cells were immunofluorescently labeled with the H3K79me3 antibody (left) diluted 1:300 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa568 or with DAPI (right). Figure B and C: staining of the cells with the H3K79me3 antibody after incubation of the antibody with 2 ng\/\u03bcl H4K20me3 and H3K79me3 peptide, respectively.
Histone extracts of HeLa cells (15 \u03bcg) were analyzed by Western blot using the Bioss antibody against H3K79me3 (Cat. No. bs-53121R), diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The molecular weight marker is shown on the right; the location of the protein of interest is indicated on the left.
A Dot Blot analysis was performed to test the cross-reactivity of the Bioss antibody against H3K79me3 (Cat. No. bs-53121R) with peptides containing other histone modifications and the unmodified H3K79. 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 Bioss antibody directed against H3K79me3 (Cat. No. bs-53121R), 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 1:3,500.

Catalog #1: bs-53121R

Applications: WB, ELISA, IF, dot-blot

Reactivity: Human, Mouse, Yeast

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 the Bioss antibody against H3K27me3 (Cat. No. bs-53122R) diluted 1:500 in TBS-Tween containing 5% skimmed milk.
A Dot Blot analysis was performed to test the cross-reactivity of the Bioss antibody against H3K27me3 (Cat. No. bs-53122R) with peptides containing other modifications of histone H3 and H4 and the unmodified H3K27 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 1:5,000. Figure 4 shows a high specificity of the antibody for the modification of interest. Please note that that antibody also recognizes the modification if S28 is phosphorylated.
Mouse NIH3T3 cells were stained with the Bioss antibody against H3K27me3 (Cat. No. bs-53122R) 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 H3K27me3 antibody (left) diluted 1:200 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 determine the titer of the antibody, an ELISA was performed using a serial dilution of H3K27me3 Polyclonal Antibody (Cat. No. bs-53122R). 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 1:3,500.

Catalog #1: bs-53122R

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

Reactivity: Human, Mouse, Rat, Others, Zebrafish, Drosophila

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 H3 (lane 3) using H3K79me1 Polyclonal Antibody (Cat. No. bs-53126R). The antibody was diluted 1:200 in TBS-Tween containing 5% skimmed milk.
A Dot Blot analysis was performed to test the cross-reactivity of the Bioss antibody against H3K79me1 (Cat. No. bs-53126R) with peptides containing other histone modifications and the unmodified H3K79. 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.
To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Bioss antibody directed against H3K79me1 (Cat. No. bs-53126R). 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 1:11,500.
ChIP assays were performed using human HeLa cells, the Bioss antibody against H3K79me1 (Cat. No. bs-53126R) and optimized PCR primer pairs for qPCR. ChIP was performed 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 a region 1 kb upstream of the promoter and the coding region of the active GAPDH gene, 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).

Catalog #1: bs-53126R

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

Reactivity: Human, Yeast

Conjugate: Unconjugated

U2OS cells were stained with the Bioss antibody against H3K36me1 (cat. No. bs-53131R) and with DAPI. Cells were fixed with 4% formaldehyde for 20\u2019 and blocked with PBS\/TX-100 containing 5% normal goat serum. Figure A: cells were immunofluorescently labeled with the H3K36me1 antibody (left) diluted 1:500 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa568 or with DAPI (right), which specifically labels DNA. Figure B, C, D, and E: staining of the cells with the H3K36me1 antibody after incubation of the antibody with 2 ng\/\u03bcl blocking peptide containing the unmodified and the mono-, di- and trimethylated H3K36, respectively.
Histone extracts (15 \u03bcg) from HeLa cells were analyzed by Western blot using the Bioss antibody against H3K36me1 (cat. No. bs-53131R) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk.
A Dot Blot analysis was performed to test the cross-reactivity of the Bioss antibody against H3K36me1 (cat. No. bs-53131R) with peptides containing other modifications of histone H3 or the unmodified H3K36. 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.
To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Bioss antibody directed against H3K36me1 (cat. No. bs-53131R), crude serum and Flow Through. 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 1:46,000.

Catalog #1: bs-53131R

Applications: WB, ELISA, IF, dot-blot

Reactivity: Human

Conjugate: Unconjugated

Western blot was performed on histone extracts (15 \u03bcg, lane 1) from HeLa cells, and on 1 \u03bcg of recombinant histone H3 (lane 2) using H3K27me3S28p Polyclonal Antibody (Cat. No. bs-53132R) diluted 1:100 in TBS-Tween containing 5% skimmed milk.
A Dot Blot analysis was performed to test the cross-reactivity of the Bioss antibody against H3K27me3S28p (Cat. No. bs-53132R) with peptides containing other modifications of histone H3 and H4 and the unmodified H3K27S28 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 1:2,000.
To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Bioss antibody directed against H3K27me3S28p (Cat. No. bs-53132R). 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 1:4,300.
ChIP assays was performed using HeLa cells treated with colcemid, H3K27me3S28p Polyclonal Antibody (Cat. No. bs-53132R) and optimized PCR primer sets for qPCR. ChIP was performed with a ChIP-seq kit, 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 gene GAPDH as a negative control and for the coding regions of the inactive genes MYT1 and TSH2B 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 H3K27me3S28p is preferably present at inactive genes.

Catalog #1: bs-53132R

Applications: WB, ELISA, dot-blot

Reactivity: Human

Conjugate: Unconjugated

HeLa cells were stained with the Bioss antibody against H3K18ac (cat. No. bs-53137R) 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 H3K18ac antibody (left) diluted 1:200 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 (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 the Bioss antibody against H3K18ac (cat. No. bs-53137R). The antibody was diluted 1:500 in TBS-Tween containing 5% skimmed milk.
To test the cross-reactivity of the Bioss antibody against H3K18ac (cat. No. bs-53137R), a Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H3K18. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:5,000. 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 against H3K18ac (cat. No. bs-53137R). 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 1:4,300.

Catalog #1: bs-53137R

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

Reactivity: Human, Mouse

Conjugate: Unconjugated

Western blot was performed on histone extracts (30 \u03bcg) from HeLa cells using H3K64me3 Polyclonal Antibody (bs-53144R). The antibody was diluted 1:100 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 H3K64me3 Polyclonal Antibody (bs-53144R) a Dot Blot was performed with peptides containing different modifications of histone H3 and H4 or the unmodified H3K64 sequence. 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 1:5,000. Figure shows a high specificity of the antibody for the modification of interest.
To determine the titer, an ELISA was performed using H3K64me3 Polyclonal Antibody (bs-53144R) 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 1:5,500.
ChIP assays were performed using K562 cells, \r\nH3K64me3 Polyclonal Antibody (bs-53144R) and optimized PCR primer sets for qPCR. ChIP was performed with 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 Sat2 and Sata satellite repeats, used as positive controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

Catalog #1: bs-53144R

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

Reactivity: Human

Conjugate: Unconjugated

To test the cross reactivity of the H3K56ac Polyclonal Antibody (bs-53145R), a Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H3K56. 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 H3K56ac Polyclonal Antibody (bs-53145R) 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 1:15,300.
ChIP assays were performed using HeLa cells, H3K56ac Polyclonal Antibody (bs-53145R) and optimized PCR primer sets for qPCR. ChIP was performed with sheared chromatin from 1.5 million cells. A titration of the antibody consisting of 0.5, 1, 2 and, 5 \u03bcg per ChIP experiment was analyzed. IgG (1 \u03bcg\/IP) was used as negative IP control. QPCR was performed with primers for a region approximately 1 kb upstream of the GAPDH promoter and for the EIF4A2 promoter, used as positive controls, and for the coding region of 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).
ChIP was performed on sheared chromatin from 1.5 million HeLaS3 cells using 5 \u03bcg of H3K56ac Polyclonal Antibody (bs-53145R) as described previously. The IP\u2019d DNA was subsequently analyzed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer\u2019s instructions. The 51 bp tags were aligned to the human genome using the BWA algorithm. Figure shows the enrichment along the complete sequence and a 1 Mb region of the X-chromosome (fig A and B) and in genomic regions of chromosome 12 and 3, surrounding the GAPDH and EIF4A2 positive control genes.

Catalog #1: bs-53145R

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

Reactivity: Human

Conjugate: Unconjugated

Catalog #1: bs-53090R

Applications: ELISA, dot-blot

Reactivity: Human

Conjugate: Unconjugated

Catalog #1: bs-53081R

Applications: ELISA

Reactivity: Human

Conjugate: Unconjugated

Catalog #1: bs-53089R

Applications: WB, ELISA

Reactivity: Human

Conjugate: Unconjugated

Catalog #1: bs-53083R

Applications: ELISA

Reactivity: 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 H2AK5ac Polyclonal Antibody (bs-53146R). The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The marker (in kDa) is shown on the left.
To test the cross reactivity of H2AK5ac Polyclonal Antibody (bs-53146R), a Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H2AK5. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:5,000. Figure shows a high specificity of the antibody for the modification of interest.
HeLa cells were stained with tH2AK5ac Polyclonal Antibody (bs-53146R) 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 H2AK5ac 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 determine the titer of the antibody, an ELISA was performed using a serial dilution of H2AK5ac Polyclonal Antibody (bs-53146R) 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 1:25,000.

Catalog #1: bs-53146R

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

Reactivity: Human

Conjugate: Unconjugated

Catalog #1: bs-53073R

Applications: WB, ELISA

Reactivity: Human

Conjugate: Unconjugated

Catalog #1: bs-53088R

Applications: ELISA

Reactivity: Human, Mouse

Conjugate: Unconjugated

Whole cell extracts (40 \u03bcg) from HeLa cells transfected with HDAC2 siRNA (lane 2) and from an untransfected control (lane 1) were analysed by Western blot using HDAC2 (7G5) Monoclonal Antibody (bsm-53026M) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right (expected size: 55 kDa); the marker (in kDa) is shown on the left.
HeLa cells were stained with HDAC2 (7G5) Monoclonal Antibody (bsm-53026M) 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 HDAC2 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.
ChIP assays were performed using HeLa cells, HDAC2 (7G5) Monoclonal Antibody (bsm-53026M) and optimized PCR primer sets for qPCR. ChIP was performed using sheared chromatin from 4 million cells. A titration of the antibody consisting of 1, 2 and 5 \u03bcg per ChIP experiment was analysed. IgG (2 \u03bcg\/IP) was used as negative IP control. QPCR was performed with primers for the ACTB, EIF4A2 and LMO4 genes, used as positive controls, and for the Sat2 satellite repeat, used as negative control. 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 on sheared chromatin from 4 million HeLa cells using 1 \u03bcg of HDAC2 (7G5) Monoclonal Antibody (Cat. No. C15200201) as described above. The IP\u2019d DNA was subsequently analyzed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer\u2019s instructions. The 51 bp tags were aligned to the mouse genome using the BWA algorithm. Figure shows the enrichment along the complete sequence and a 2 Mb region of human chromosome 3 (fig 2A and B), and in a two genomic regions surrounding the LMO4 positive control gene and the MAGEC1 gene (figure 2C and D).

Catalog #1: bsm-53026M

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

Reactivity: Human

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 H2Apan Polyclonal Antibody (bs-53140R). The antibody was diluted 1:2,000 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 H2Apan Polyclonal Antibody (bs-53140R) 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 H2Apan 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 determine the titer of the antibody, an ELISA was performed using a serial dilution of H2Apan Polyclonal Antibody (bs-53140R) in antigen-coated wells. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:32,500.
ChIP assays were performed using HeLa cells, \r\nH2Apan Polyclonal Antibody (bs-53140R) and optimized PCR primer sets for qPCR. ChIP was performed with sheared chromatin from 1 million cells using the IP-Star automated system. 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 with primers for the GAPDH and EIF4A2 promoters, used as negative controls and for the inactive MYOD1 gene and the Sat2 satellite repeat, used 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).

Catalog #1: bs-53140R

Applications: WB, ELISA, IF(ICC)

Reactivity: Human

Conjugate: Unconjugated

Western blot was performed on nuclear extracts from mouse fibroblastst (NIH3T3) and embryonic stem cells (E14Tg2a) with the antibody against MBD2 (bs-53058R), diluted 1:500 in BSA\/PBS- Tween. The molecular weight marker (M, in kDa) is shown on the left; the location of the protein of interest is indicated on the right.
To determine the titer, an ELISA was performed using a serial dilution of the antibody directed against MBD2 (bs-53058R). By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:185,000.

Catalog #1: bs-53058R

Applications: WB, ELISA

Reactivity: Mouse

Conjugate: Unconjugated

Catalog #1: bs-53067R

Applications: ELISA

Reactivity: Human, Mouse

Conjugate: Unconjugated

Catalog #1: bs-53079R

Applications: ELISA

Reactivity: Human

Conjugate: Unconjugated

Western blot was performed on 150 \u03bcg nuclear extract from either NIH3T3 or HeLa cells with the TIP5 antibody (bs-53052R), diluted 1:1,000 in PBS containing 5% milk powder and 0.1% Tween- 20. The molecular weight marker is shown on the left, the location of the protein of interest is indicated on the right.
ChIP assays were performed using the TIP5 antibody (bs-53052R). Chromatin from HEK293T and NIH3T3 cells were formaldehyde cross-linked and sheared to yield fragments with an average length of 200 to 400 bp. ChIP was performed overnight at 4\u00b0C with 100 \u03bcg sheared chromatin and either 5 \u03bcl of the TIP5 antibody or 5 \u03bcl IgG which was used as negative IP control. The IP\u2019d DNA was analyzed by qPCR with primer sets for the promoter and the coding region of the 28s ribosomal RNA gene. The figure shows the recovery by the TIP5 antibody and by IgG (set to 1), normalized to the input DNA. These results show that, both in HEK293T and in NIH3T3 cells, TIP5 is associated with the promoter, but not with the coding region of the 28srRNA gene.

Catalog #1: bs-53052R

Applications: WB, ELISA

Reactivity: Human, Mouse

Conjugate: Unconjugated

Western blot was performed on nuclear extracts from HeLa cells (40 \u03bcg) with the antibody against PADI4 (bs-53047R), diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The molecular weight marker (in kDa) is shown on the left; the location of the protein of interest is indicated on the right.
To determine the titer of the antibody, an ELISA was performed using a serial dilution of antibody directed against PADI4 (bs-53047R). The wells were coated with the peptides. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1: 40,200.

Catalog #1: bs-53047R

Applications: WB, ELISA

Reactivity: Human

Conjugate: Unconjugated

Catalog #1: bs-53082R

Applications: WB, ELISA

Reactivity: Human

Conjugate: Unconjugated

Catalog #1: bs-53086R

Applications: ELISA

Reactivity: Human, Mouse

Conjugate: Unconjugated

To determine the titer of the antibody, an ELISA was performed using a serial dilution of Bioss antibody directed against human DNMT3A (Cat. No. bs-53128R), 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 1:500.
Human embryonic kidney cells (HEK293T) were transiently transfected with an expression vector for GAL4- tagged DNMT3A. Whole cell extracts were immunoprecipitated with 2 \u03bcg of DNMT3A Polyclonal Antibody (Cat. No. bs-53128R). The presence of GAL4-DNMT3A in the non-treated cell extracts and in the immunoprecipitates was demonstrated by western blot analysis with the anti-GAL4 antibody (lane 1 and 2 respectively). Alternatively, GAL4-DNMT3A was immunoprecipitated and western blot analysis was performed with the DNMT3A antibody (diluted 1:2,000) (lane 3).

Catalog #1: bs-53128R

Applications: WB, ELISA, IP, ChIP-seq

Reactivity: Human

Conjugate: Unconjugated

Catalog #1: bs-10248R-HRP

Applications: WB, ELISA, IHC-P, IHC-F

Reactivity: Mouse, Rat

Conjugate: HRP

Catalog #1: bs-10248R-Biotin

Applications: WB, ELISA, IHC-P, IHC-F

Reactivity: Mouse, Rat

Conjugate: Biotin

Mouse muscle lysates probed with CRHR1 Polyclonal Antibody, Unconjugated (bs-10248R) at 1:300 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:10000 for 60 min at 37˚C.
Mouse kidney lysates probed with CRHR1 Polyclonal Antibody, Unconjugated (bs-10248R) at 1:300 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:10000 for 60 min at 37˚C.
Paraformaldehyde-fixed, paraffin embedded Mouse brain; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with CRHR1 Polyclonal Antibody, Unconjugated (bs-10248R) at 1:200 overnight at 4\u00b0C, DAB staining.
Paraformaldehyde-fixed, paraffin embedded Rat brain; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37\u00b0C for 30min; Antibody incubation with CRHR1 Polyclonal Antibody, Unconjugated (bs-10248R) at 1:200 overnight at 4\u00b0C, DAB staining.

Catalog #1: bs-10248R

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

Reactivity: Mouse, Rat

Conjugate: Unconjugated

Catalog #1: bs-10442R-HRP

Applications: WB, ELISA, IHC-P, IHC-F

Reactivity: Human, Mouse, Rat

Conjugate: HRP

Catalog #1: bs-10442R-Biotin

Applications: WB, ELISA, IHC-P, IHC-F

Reactivity: Human, Mouse, Rat

Conjugate: Biotin

Rat lung lysates probed with Caspase-1 p20 Polyclonal Antibody, Unconjugated (bs-10442R) at 1:300 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:10000 for 60 min at 37˚C.
HL-60 cells were fixed with 4% PFA for 10min at room temperature,permeabilized with 0.1% PBST for 20 min at room temperature, and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with Caspase-1 p20 Polyclonal Antibody(bs-10442R)at 1:100 dilution in blocking buffer and incubated for 30 min at room temperature, washed twice with 2%BSA in PBS, followed by secondary antibody incubation for 40 min at room temperature. Acquisitions of 20,000 events were performed. Cells stained with primary antibody (green), and isotype control (orange).
Lane 1: Mouse Heart lysates; Lane 2: Rat Heart lysates probed with Caspase-1 p20 Polyclonal Antibody, Unconjugated (bs-10442R) at 1:1000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.

Catalog #1: bs-10442R

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

Reactivity: Human, Mouse, Rat

Conjugate: Unconjugated

Catalog #1: bs-10551R-Biotin

Applications: WB, ELISA

Reactivity: Human

Conjugate: Biotin

Catalog #1: bs-10551R-HRP

Applications: WB, ELISA

Reactivity: Human

Conjugate: HRP

Catalog #1: bs-10557R

Applications: WB, ELISA

Reactivity: Human, Mouse

Conjugate: Unconjugated

Catalog #1: bs-10551R

Applications: WB, ELISA, FCM

Reactivity: Human

Conjugate: Unconjugated

Catalog #1: bs-10557R-Biotin

Applications: WB, ELISA

Reactivity: Human, Mouse

Conjugate: Biotin

Catalog #1: bs-10557R-HRP

Applications: WB, ELISA

Reactivity: Human, Mouse

Conjugate: HRP

Catalog #1: bs-10576R-Biotin

Applications: WB, ELISA, IHC-P, IHC-F

Reactivity: Human

Conjugate: Biotin

Catalog #1: bs-10576R-HRP

Applications: WB, ELISA, IHC-P, IHC-F

Reactivity: Human

Conjugate: HRP

Catalog #1: bs-10576R

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

Reactivity: Human

Conjugate: Unconjugated

Catalog #1: bs-10592R

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

Reactivity: Human

Conjugate: Unconjugated

Catalog #1: bs-10592R-HRP

Applications: WB, ELISA, IHC-P, IHC-F

Reactivity: Human

Conjugate: HRP

Catalog #1: bs-10603R-Biotin

Applications: WB, ELISA, IHC-P, IHC-F

Reactivity: Mouse

Conjugate: Biotin