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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

Nuclear extracts from HeLa cells (40 \u03bcg) were analyzed by Western blot using the Bioss antibody against JMJD2a (Cat. No. bs-53136R) diluted 1:500 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right (expected size 121 kDa); the marker (in kDa) is shown on the left.
To determine the titer of the antibody, an ELISA was performed using a serial dilution of JMJD2a Polyclonal Antibody (bs-53136R), crude serum and flow through. The plates were coated with the peptide used for immunization of the rabbit. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:9,000.

Catalog #1: bs-53136R

Applications: WB, ELISA, IF

Reactivity: Human

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

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 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

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

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

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