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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

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

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

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-53082R

Applications: WB, ELISA

Reactivity: Human

Conjugate: Unconjugated

Catalog #1: bs-53079R

Applications: ELISA

Reactivity: Human

Conjugate: Unconjugated

Catalog #1: bs-53078R

Applications: ELISA

Reactivity: Human, Mouse

Conjugate: Unconjugated

Catalog #1: bs-53073R

Applications: WB, ELISA

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

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

Catalog #1: bs-53067R

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

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

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

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

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

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

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 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

Catalog #1: bs-53089R

Applications: WB, ELISA

Reactivity: Human

Conjugate: Unconjugated

Catalog #1: bs-53081R

Applications: ELISA

Reactivity: Human

Conjugate: Unconjugated

Catalog #1: bs-53077R

Applications: ELISA

Reactivity: Human

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

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

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

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 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

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

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

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 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

Catalog #1: bs-53090R

Applications: ELISA, dot-blot

Reactivity: Human

Conjugate: Unconjugated

Catalog #1: bs-53088R

Applications: ELISA

Reactivity: Human, Mouse

Conjugate: Unconjugated

Catalog #1: bs-53086R

Applications: ELISA

Reactivity: Human, Mouse

Conjugate: Unconjugated

Catalog #1: bs-53085R

Applications: WB, ELISA

Reactivity: Human, Mouse

Conjugate: Unconjugated

Catalog #1: bs-53084R

Applications: WB, ELISA

Reactivity: Mouse

Conjugate: Unconjugated

Catalog #1: bs-53083R

Applications: ELISA

Reactivity: Mouse

Conjugate: Unconjugated

Catalog #1: bs-53080R

Applications: WB, ELISA

Reactivity: Human

Conjugate: Unconjugated

To determine the titer, an ELISA was performed using a serial dilution of the EHMT1 antibody (bs-53068R). The wells were coated with peptides. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:2,000.
Nuclear extracts of HeLa cells (40 \u03bcg) were analyzed by Western blot using the EHMT1 antibody (bs-53068R) 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 smaller fragment may represent a splicing variant.

Catalog #1: bs-53068R

Applications: WB, ELISA, IHC-P

Reactivity: Human, Mouse

Conjugate: Unconjugated

Catalog #1: bs-53057R

Applications: ELISA

Reactivity: Mouse

Conjugate: Unconjugated

Catalog #1: bs-53056R

Applications: ELISA

Reactivity: Mouse

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 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

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

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 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

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