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Catalog #1: bs-51111r-50ug

Applications: WB, ELISA, IF, dot-blot

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

Catalog #1: bs-51121r-50ug

Applications: WB, ELISA, dot-blot

Reactivity: Human, Mouse, Yeast

Conjugate: Unconjugated

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

Catalog #1: bs-53093R

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

Reactivity: Human, Mouse, Others

Conjugate: Unconjugated

Catalog #1: bs-51093r-50ug

Applications: WB, ELISA, IF, dot-blot

Reactivity: Human, Mouse, Others

Conjugate: Unconjugated

Catalog #1: bs-51022R

Applications: WB, ELISA, IF

Reactivity: Human

Conjugate: Unconjugated

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

Catalog #1: bs-53092R

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

Reactivity: Human, Mouse, Others

Conjugate: Unconjugated

Catalog #1: bs-51092r-50ug

Applications: WB, ELISA, IF, dot-blot

Reactivity: Human, Mouse, 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

Catalog #1: bs-51062r-100ul

Applications: WB, ELISA, IF, dot-blot

Reactivity: Human

Conjugate: Unconjugated

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

Catalog #1: bs-53043R

Applications: WB, ELISA, IF, dot-blot

Reactivity: Human

Conjugate: Unconjugated

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

Catalog #1: bs-51043r-100ul

Applications: WB, ELISA, IF, dot-blot

Reactivity: Human

Conjugate: Unconjugated

Catalog #1: bs-51120r-50ug

Applications: WB, ELISA, dot-blot

Reactivity: Human

Conjugate: Unconjugated

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

Catalog #1: bsm-53016M

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

Reactivity: Human

Conjugate: Unconjugated

Catalog #1: bs-51016R

Applications: WB, ELISA, IF

Reactivity: Human

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

Catalog #1: bs-51118r-50ug

Applications: WB, ELISA, IF, dot-blot

Reactivity: Human, Mouse, Others

Conjugate: Unconjugated

Catalog #1: bs-53029R

Applications: WB

Reactivity: Human, Mouse

Conjugate: Unconjugated

Catalog #1: bs-51029r-100ul

Applications: WB

Reactivity: Human, Mouse, Drosophila

Conjugate: Unconjugated

Catalog #1: bs-51114r-50ug

Applications: WB, ELISA, IF, dot-blot

Reactivity: Human, Mouse, Others, Zebrafish

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

Catalog #1: bs-51072r-100ul

Applications: WB, ELISA, IF, dot-blot

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 H3K9un (7D7) Monoclonal Antibody (bsm-53024M). The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.
HeLa cells were stained with H3K9un (7D7) Monoclonal Antibody (bsm-53024M) 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 H3K9un 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, H3K9un (7D7) Monoclonal Antibody (bsm-53024M) and optimized PCR primer sets for qPCR. ChIP was performed on 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 analysed. IgG (2 \u03bcg\/IP) was used as negative IP control. QPCR was performed with primers for the coding regions of the MYOD1 and MB genes and for a region 1 kb upstream of the GAPDH promoter, used as positive controls, and for the ZNF510 coding region, the EIF4A2 promoter and the Sat2 satellite repeat region, used as negative controls. Figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

Catalog #1: bsm-53024M

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

Reactivity: Human

Conjugate: Unconjugated

Catalog #1: bs-10320R

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

Reactivity: Virus

Conjugate: Unconjugated

Western blot was performed on whole cell extracts from HeLa cells (25 \u03bcg, lane 1) and recombinant histone H2A, H2B, H3 and H4 (1ug, lane 2, 3, 4 and 5, respectively) using the H3pan (1B2) Monoclonal Antibody (bsm-53008M). The antibody was diluted 1:1,000 in TBSTween containing 5% skimmed milk, followed by secondary antibody incubation and exposure.
Western blot was performed on whole cell extracts (30\u03bcg) from different cell types (lane 1: HeLa, lane 2: K562, lane 3: MCF7, lane 4: U2OS, lane 5: HepG2, lane 6: Jurkat, lane 7: NIH3T3, lane 8: E14Tg2a mouse ES cells) using H3pan Monoclonal Antibody (bsm-53008M) at 1:1,000 dilution in TBS-Tween containing 5% skimmed milk, followed by secondary antibody incubation and exposure.
HeLa cells stained with the H3pan Monoclonal Antibody (bsm-53008M) and with DAPI. Cells were fixed with 4% formaldehyde for 10\u2019 and blocked with PBS\/TX-100 containing 1% BSA. The cells were labeled with the H3 antibody (middle) diluted 1:500 in blocking solution followed by an anti-mouse antibody conjugated to Alexa594. The left panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.

Catalog #1: bsm-53008M

Applications: WB, IF(ICC)

Reactivity: Human, Others

Conjugate: Unconjugated

Catalog #1: bs-51008r-50ug

Applications: WB

Reactivity: Human, Others

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

Catalog #1: bs-51074r-100ul

Applications: WB, ELISA, dot-blot

Reactivity: Human, Mouse, Rat

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

Catalog #1: bs-51054r-100ul

Applications: WB, ELISA, dot-blot

Reactivity: Human

Conjugate: Unconjugated

Catalog #1: bs-51141r-50ug

Applications: WB, ELISA, IF, dot-blot

Reactivity: Human, Mouse

Conjugate: Unconjugated

Catalog #1: bs-53141R

Applications: WB, ELISA, IF, dot-blot

Reactivity: Human, Mouse

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

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

Catalog #1: bs-51069r-100ul

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

Reactivity: Human

Conjugate: Unconjugated

Catalog #1: bs-51135r-50ug

Applications: WB, ELISA, IF, dot-blot

Reactivity: Human

Conjugate: Unconjugated

Catalog #1: bs-95113c-1ml10mmin-dmso

Applications: Life Science

Reactivity:

Conjugate: Unconjugated

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

Catalog #1: bs-53035R

Applications: WB, ELISA, dot-blot

Reactivity: Human, Others

Conjugate: Unconjugated

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

Catalog #1: bs-51035r-100ul

Applications: WB, ELISA, dot-blot

Reactivity: Human

Conjugate: Unconjugated

Catalog #1: bs-51104r-50ug

Applications: WB, ELISA, dot-blot

Reactivity: Human

Conjugate: Unconjugated

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

Catalog #1: bs-53115R

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

Reactivity: Human, Mouse, Others

Conjugate: Unconjugated

Catalog #1: bs-51115r-50ug

Applications: WB, ELISA, IF, dot-blot

Reactivity: Human, Mouse

Conjugate: Unconjugated

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

Catalog #1: bs-53098R

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

Reactivity: Human, Mouse

Conjugate: Unconjugated

Catalog #1: bs-51098r-50ug

Applications: ELISA, IF, dot-blot

Reactivity: Human, Mouse

Conjugate: Unconjugated

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

Catalog #1: bs-53096R

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

Reactivity: Human, Mouse

Conjugate: Unconjugated

Catalog #1: bs-51096r-50ug

Applications: WB, ELISA, IF, dot-blot

Reactivity: Human, Mouse

Conjugate: Unconjugated