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

Catalog #1: bs-53109R

Applications: WB, ELISA, IF, dot-blot

Reactivity: Human

Conjugate: Unconjugated

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

Catalog #1: bs-53108R

Applications: WB, ELISA, IF, dot-blot

Reactivity: Human, Others

Conjugate: Unconjugated

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

Catalog #1: bs-53106R

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

Reactivity: Human, Mouse

Conjugate: Unconjugated

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

Catalog #1: bs-53105R

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

Reactivity: Human

Conjugate: Unconjugated

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

HeLa cells were stained with the Bioss antibody against H3K4me3 (Cat. No. bs-53103R) 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 H3K4me3 antibody (left) diluted 1:100 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 (40 \u00b5g, lane 1) and histone extracts (15 \u00b5g, lane 2) from HeLa cells, and on 1 \u00b5g of recombinant histone H2A, H2B, H3.1, H3.3 and H4 (lane 3, 4, 5, 6 and 7, respectively) using the Bioss antibody against H3K4me3 (Cat. No. bs-53103R). The antibody was diluted 1:500 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is shown 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 H3K4me3 (Cat. No. bs-53103R) with peptides containing other modifications and unmodified sequences of histone H3 and H4. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:2,000. The figure shows a high specificity of the antibody for the modification of interest.
ChIP was performed on sheared chromatin from 1 million HeLa cells using 1 \u00b5g of the Bioss antibody against H3K4me3 (Cat. No. bs-53103R) as described above. The IP'd DNA was subsequently analyzed on an Illumina Genome Analyzer. Library preparation, cluster generation, and sequencing were performed according to the manufacturer's instructions. The 50 bp tags were aligned to the human genome using the BWA algorithm. The figure shows the peak distribution along the complete sequence and a 1.2 Mb region of the X-chromosome (figure A and B) and in two regions surrounding the GAPDH and EIF4A2 positive control genes, respectively (figure C and D). These results clearly show an enrichment of the H3K4 trimethylation at the promoters of active genes.

Catalog #1: bs-53103R

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

Reactivity: Human, Mouse, Others

Conjugate: Unconjugated

HeLa cells were stained with the Bioss antibody against WDR5 (Cat. No. bs-53101R) 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 WDR5 antibody (left) diluted 1:1,000 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.
Nuclear extracts from HeLa cells (20 \u03bcg) were analyzed by Western blot using the Bioss antibody against WDR5 (Cat. No. bs-53101R) 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.
ChIP was performed on sheared chromatin from 4 million HeLa cells using 2 \u03bcg of the Bioss antibody against WDR5 (Cat. No. bs-53101R) 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 50 bp tags were aligned to the human genome using the BWA algorithm. The figure shows the enrichment along the complete sequence and a 2 Mb region of human chromosome 1 (fig A and B), and in two genomic regions surrounding the RPL41 and RPS4X positive control genes (fig C and D).
ChIP assays were performed using HeLa cells, the Bioss antibody against WDR5 (Cat. No. bs-53101R) and optimized PCR primer sets for qPCR. ChIP was performed with a ChIP-seq kit, using sheared chromatin from 4 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 optimized primers for the promoters of the RPL41 and RPS4X ribosomal protein genes, used as positive controls, and for the MYOD1 gene and the Sat2 satellite repeat, used as negative controls. The figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

Catalog #1: bs-53101R

Applications: WB, IF, ChIP-seq

Reactivity: Human

Conjugate: Unconjugated

Immunoprecipitation was performed on nuclear extracts from cells transfected with FLAG-tagged L3MBTL1 (L3MBTL1-F) with the Bioss antibody against L3MBTL1 (Cat. No. bs-53097R) and with an IgG negative control antibody. Western Blot was performed with an anti-FLAG antibody to demonstrate the presence of L3MBTL1-F in the input (lane 1), in the precipitated fraction (P) and in the supernatant (SN). These data show that L3MBTL1-F was efficiently precipitated by the L3MBTL1 antibody (lane 3 and 5), whereas it was not precipitated by the IgG negative control antibody (lane 2 and 4).
Western blot was performed on nuclear extracts from HeLa cells (HeLa NE) using the Bioss antibody against L3MBTL1 (Cat. No. bs-53097R) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk (lane 2). A molecular weight marker is shown in lane 1; the location of the protein of interest is indicated on the right.

Catalog #1: bs-53097R

Applications: WB, IP, IF

Reactivity: Human

Conjugate: Unconjugated

Western blot was performed on HeLa nuclear extracts (lane 1) and purified human SKI complex (lane 2) (Figure A), or on nuclear extracts from HeLa cells (HeLa NE, 20 \u03bcg) (Figure B) using the Bioss antibody against SKI3 (Cat. No. bs-53094R) diluted 1:1,000.
HeLa cells were stained with the Bioss antibody against SKI3 (Cat. No. bs-53094R) (red), with DAPI to label the DNA (blue), and with an antibody against \u03b1-tubulin to label the cytoplasm (green). SKI3 is detected in both the cytoplasm and the nucleus but appears to be excluded from the nucleolus.

Catalog #1: bs-53094R

Applications: WB, IF(ICC), IF

Reactivity: Human

Conjugate: Unconjugated

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

Catalog #1: bs-53136R

Applications: WB, ELISA, IF

Reactivity: Human

Conjugate: Unconjugated

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

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

Catalog #1: bs-53133R

Applications: WB, ELISA, IF, dot-blot

Reactivity: Human, Mouse

Conjugate: Unconjugated

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

Catalog #1: bs-53134R

Applications: WB, ELISA

Reactivity: Human

Conjugate: Unconjugated

Catalog #1: bsm-53018M

Applications: WB

Reactivity: Human, Mouse

Conjugate: Unconjugated

Whole cell extracts from HL60 and NB4 cells (40 \u03bcg) were analysed by Western blot using TET2 (6A1) Monoclonal Antibody (bsm-53017M), diluted 1:2,000 in PBS containing 10% milk. The position of the protein of interest (expected MW 224 kDa) is indicated on the right; the marker (in kDa) is shown on the left.
IP was performed on 250 \u03bcg HL60 RIPA cell lysate using TET2 (6A1) Monoclonal Antibody (bsm-53017M) (lane 3) or an IgG negative control (lane 2). The input sample (25 \u03bcg RIPA lysate) was used as a positive control (lane 1). The samples were analysed by Western blot using TET2 (6A1) Monoclonal Antibody (bsm-53017M), diluted 1:2,000 in PBS containing 10% milk.

Catalog #1: bsm-53017M

Applications: WB, IP

Reactivity: Human, Mouse

Conjugate: Unconjugated

Western blot was performed on whole cell (25 \u03bcg, lane 1) and histone extracts (15 \u03bcg, lane 2) from HeLa cells, and on 1 \u03bcg of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using H2AK5ac Polyclonal Antibody (bs-53146R). The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The marker (in kDa) is shown on the left.
To test the cross reactivity of H2AK5ac Polyclonal Antibody (bs-53146R), a Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H2AK5. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:5,000. Figure shows a high specificity of the antibody for the modification of interest.
HeLa cells were stained with tH2AK5ac Polyclonal Antibody (bs-53146R) and with DAPI. Cells were fixed with 4% formaldehyde for 10\u2019 and blocked with PBS\/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labeled with the H2AK5ac antibody (left) diluted 1:500 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.
To determine the titer of the antibody, an ELISA was performed using a serial dilution of H2AK5ac Polyclonal Antibody (bs-53146R) in antigen-coated wells. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:25,000.

Catalog #1: bs-53146R

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

Reactivity: Human

Conjugate: Unconjugated

To test the cross reactivity of the H3K56ac Polyclonal Antibody (bs-53145R), a Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H3K56. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure shows a high specificity of the antibody for the modification of interest.
To determine the titer of the antibody, an ELISA was performed using a serial dilution of H3K56ac Polyclonal Antibody (bs-53145R) in antigen-coated wells. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:15,300.
ChIP assays were performed using HeLa cells, H3K56ac Polyclonal Antibody (bs-53145R) and optimized PCR primer sets for qPCR. ChIP was performed with sheared chromatin from 1.5 million cells. A titration of the antibody consisting of 0.5, 1, 2 and, 5 \u03bcg per ChIP experiment was analyzed. IgG (1 \u03bcg\/IP) was used as negative IP control. QPCR was performed with primers for a region approximately 1 kb upstream of the GAPDH promoter and for the EIF4A2 promoter, used as positive controls, and for the coding region of the inactive MYOD1 gene and the Sat2 satellite repeat, used as negative controls. Figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).
ChIP was performed on sheared chromatin from 1.5 million HeLaS3 cells using 5 \u03bcg of H3K56ac Polyclonal Antibody (bs-53145R) as described previously. The IP\u2019d DNA was subsequently analyzed on an Illumina HiSeq. Library preparation, cluster generation and sequencing were performed according to the manufacturer\u2019s instructions. The 51 bp tags were aligned to the human genome using the BWA algorithm. Figure shows the enrichment along the complete sequence and a 1 Mb region of the X-chromosome (fig A and B) and in genomic regions of chromosome 12 and 3, surrounding the GAPDH and EIF4A2 positive control genes.

Catalog #1: bs-53145R

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

Reactivity: Human

Conjugate: Unconjugated

Western blot was performed on histone extracts (30 \u03bcg) from HeLa cells using H3K64me3 Polyclonal Antibody (bs-53144R). The antibody was diluted 1:100 in TBS-Tween containing 5% skimmed milk. The marker (in kDa) is shown on the left, the position of the protein is indicated on the right.
To check the specificity of H3K64me3 Polyclonal Antibody (bs-53144R) a Dot Blot was performed with peptides containing different modifications of histone H3 and H4 or the unmodified H3K64 sequence. One hundred to 0.2 pmol of peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:5,000. Figure shows a high specificity of the antibody for the modification of interest.
To determine the titer, an ELISA was performed using H3K64me3 Polyclonal Antibody (bs-53144R) in antigen coated wells. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:5,500.
ChIP assays were performed using K562 cells, \r\nH3K64me3 Polyclonal Antibody (bs-53144R) and optimized PCR primer sets for qPCR. ChIP was performed with sheared chromatin from 1 million cells. A titration of the antibody consisting of 1, 2, 5, and 10 \u03bcg per ChIP experiment was analyzed. IgG (2 \u03bcg\/IP) was used as negative IP control. QPCR was performed with primers for the promoter of the active GAPDH and EIF4A2 genes, used as negative controls, and for the Sat2 and Sata satellite repeats, used as positive controls. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

Catalog #1: bs-53144R

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

Reactivity: Human

Conjugate: Unconjugated

Western blot was performed on whole cell (25 \u03bcg, lane 1) and histone extracts (15 \u03bcg, lane 2) from HeLa cells, and on 1 \u03bcg of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using H2Apan Polyclonal Antibody (bs-53140R). The antibody was diluted 1:2,000 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the right; the marker (in kDa) is shown on the left.
HeLa cells were stained with H2Apan Polyclonal Antibody (bs-53140R) and with DAPI. Cells were fixed with 4% formaldehyde for 10\u2019 and blocked with PBS\/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labeled with the H2Apan antibody (left) diluted 1:500 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.
To determine the titer of the antibody, an ELISA was performed using a serial dilution of H2Apan Polyclonal Antibody (bs-53140R) in antigen-coated wells. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:32,500.
ChIP assays were performed using HeLa cells, \r\nH2Apan Polyclonal Antibody (bs-53140R) and optimized PCR primer sets for qPCR. ChIP was performed with sheared chromatin from 1 million cells using the IP-Star automated system. A titration of the antibody consisting of 1, 2, 5, and 10 \u03bcg per ChIP experiment was analyzed. IgG (5 \u03bcg\/IP) was used as negative IP control. QPCR was performed with primers for the GAPDH and EIF4A2 promoters, used as negative controls and for the inactive MYOD1 gene and the Sat2 satellite repeat, used as positive controls. Figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

Catalog #1: bs-53140R

Applications: WB, ELISA, IF(ICC)

Reactivity: Human

Conjugate: Unconjugated

Western blot was performed on histone extracts (15 \u03bcg, lane 1) from HeLa cells, and on 1 \u03bcg of recombinant histone H3 (lane 2) using H3K27me3S28p Polyclonal Antibody (Cat. No. bs-53132R) diluted 1:100 in TBS-Tween containing 5% skimmed milk.
A Dot Blot analysis was performed to test the cross-reactivity of the Bioss antibody against H3K27me3S28p (Cat. No. bs-53132R) with peptides containing other modifications of histone H3 and H4 and the unmodified H3K27S28 sequence. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:2,000.
To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Bioss antibody directed against H3K27me3S28p (Cat. No. bs-53132R). The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:4,300.
ChIP assays was performed using HeLa cells treated with colcemid, H3K27me3S28p Polyclonal Antibody (Cat. No. bs-53132R) and optimized PCR primer sets for qPCR. ChIP was performed with a ChIP-seq kit, using sheared chromatin from 1 million cells. A titration of the antibody consisting of 1, 2, 5, and 10 \u03bcg per ChIP experiment was analyzed. IgG (2 \u03bcg\/IP) was used as negative IP control. QPCR was performed with primers for the promoter of the active gene GAPDH as a negative control and for the coding regions of the inactive genes MYT1 and TSH2B as positive controls. The figure shows the recovery expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis). These results are in accordance with the observation that H3K27me3S28p is preferably present at inactive genes.

Catalog #1: bs-53132R

Applications: WB, ELISA, dot-blot

Reactivity: Human

Conjugate: Unconjugated

U2OS cells were stained with the Bioss antibody against H3K36me1 (cat. No. bs-53131R) and with DAPI. Cells were fixed with 4% formaldehyde for 20\u2019 and blocked with PBS\/TX-100 containing 5% normal goat serum. Figure A: cells were immunofluorescently labeled with the H3K36me1 antibody (left) diluted 1:500 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa568 or with DAPI (right), which specifically labels DNA. Figure B, C, D, and E: staining of the cells with the H3K36me1 antibody after incubation of the antibody with 2 ng\/\u03bcl blocking peptide containing the unmodified and the mono-, di- and trimethylated H3K36, respectively.
Histone extracts (15 \u03bcg) from HeLa cells were analyzed by Western blot using the Bioss antibody against H3K36me1 (cat. No. bs-53131R) diluted 1:1,000 in TBS-Tween containing 5% skimmed milk.
A Dot Blot analysis was performed to test the cross-reactivity of the Bioss antibody against H3K36me1 (cat. No. bs-53131R) with peptides containing other modifications of histone H3 or the unmodified H3K36. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000.
To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Bioss antibody directed against H3K36me1 (cat. No. bs-53131R), crude serum and Flow Through. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:46,000.

Catalog #1: bs-53131R

Applications: WB, ELISA, IF, dot-blot

Reactivity: Human

Conjugate: Unconjugated

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

HeLa cells were stained with the Bioss antibody against HP1\u03b1 (Cat. No. bs-53123R) 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 HP1\u03b1 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 nuclear extracts from HeLa cells (20 \u03bcg) with the Bioss antibody against human HP1\u03b1 (Cat. No. bs-53123R), diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The molecular weight marker (in kDa) is shown on the left; the expected location of HP1\u03b1, HP1\u03b2, and HP1\u03b3 is indicated on the right.

Catalog #1: bs-53123R

Applications: WB, IF

Reactivity: Human

Conjugate: Unconjugated

Human osteosarcoma (U2OS) cells were stained with the Bioss antibody against H3K79me3 (Cat. No. bs-53121R) and with DAPI. Cells were fixed with 2.5% formaldehyde for 30\u2019 and blocked with PBS\/TX-100 containing 1% BSA. Figure A: cells were immunofluorescently labeled with the H3K79me3 antibody (left) diluted 1:300 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa568 or with DAPI (right). Figure B and C: staining of the cells with the H3K79me3 antibody after incubation of the antibody with 2 ng\/\u03bcl H4K20me3 and H3K79me3 peptide, respectively.
Histone extracts of HeLa cells (15 \u03bcg) were analyzed by Western blot using the Bioss antibody against H3K79me3 (Cat. No. bs-53121R), diluted 1:1,000 in TBS-Tween containing 5% skimmed milk. The molecular weight marker is shown on the right; the location of the protein of interest is indicated on the left.
A Dot Blot analysis was performed to test the cross-reactivity of the Bioss antibody against H3K79me3 (Cat. No. bs-53121R) with peptides containing other histone modifications and the unmodified H3K79. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:20,000.The figure shows a high specificity of the antibody for the modification of interest.
To determine the titer of the antibody, an ELISA was performed using a serial dilution of Bioss antibody directed against H3K79me3 (Cat. No. bs-53121R), crude serum and flow through in antigen-coated wells. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:3,500.

Catalog #1: bs-53121R

Applications: WB, ELISA, IF, dot-blot

Reactivity: Human, Mouse, Yeast

Conjugate: Unconjugated

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

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