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Catalog #1: bs-2001R-HRP

Applications: ELISA

Reactivity: Virus

Conjugate: HRP

Catalog #1: bs-2003R

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

Reactivity: Virus, Others

Conjugate: Unconjugated

Catalog #1: bs-15399R

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

Reactivity:

Conjugate: Unconjugated

Catalog #1: bs-15399R-Biotin

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

Reactivity:

Conjugate: Biotin

Catalog #1: bs-15399R-HRP

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

Reactivity:

Conjugate: HRP

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

Catalog #1: bs-53146R

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

Reactivity: Human

Conjugate: Unconjugated

Catalog #1: bs-51146R

Applications: WB, ELISA, 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 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

Catalog #1: bs-51140R

Applications: WB, ELISA

Reactivity: Human

Conjugate: Unconjugated

Catalog #1: bs-15401R

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

Reactivity:

Conjugate: Unconjugated

Catalog #1: bs-15401R-Biotin

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

Reactivity:

Conjugate: Biotin

Catalog #1: bs-15401R-HRP

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

Reactivity:

Conjugate: HRP

Catalog #1: bs-51023R

Applications: WB, ELISA, IF

Reactivity: Human

Conjugate: Unconjugated

Catalog #1: bsm-53023M

Applications: WB, ELISA, IF

Reactivity: Human

Conjugate: Unconjugated

Catalog #1: bs-1690R

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

Reactivity: Virus

Conjugate: Unconjugated

Catalog #1: bs-1690R-Biotin

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

Reactivity: Virus

Conjugate: Biotin

Catalog #1: bs-1690R-HRP

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

Reactivity: Virus

Conjugate: HRP

Catalog #1: bs-6191R

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

Reactivity: Human

Conjugate: Unconjugated

Catalog #1: bs-6191R-Biotin

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

Reactivity: Human

Conjugate: Biotin

Catalog #1: bs-6191R-HRP

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

Reactivity: Human

Conjugate: HRP

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

HeLa cells were stained with the Bioss antibody against H3K18ac (cat. No. bs-53137R) and with DAPI. Cells were fixed with 4% formaldehyde for 10\u2019 and blocked with PBS\/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labeled with the H3K18ac antibody (left) diluted 1:200 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.
Western blot was performed on whole cell (25 \u03bcg, lane 1) and histone extracts (15 \u03bcg, lane 2) from HeLa cells, and on 1 \u03bcg of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the Bioss antibody against H3K18ac (cat. No. bs-53137R). The antibody was diluted 1:500 in TBS-Tween containing 5% skimmed milk.
To test the cross-reactivity of the Bioss antibody against H3K18ac (cat. No. bs-53137R), a Dot Blot analysis was performed with peptides containing other histone modifications and the unmodified H3K18. One hundred to 0.2 pmol of the respective peptides were spotted on a membrane. The antibody was used at a dilution of 1:5,000. The figure shows a high specificity of the antibody for the modification of interest.
To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Bioss antibody against H3K18ac (cat. No. bs-53137R). The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:4,300.

Catalog #1: bs-53137R

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

Reactivity: Human, Mouse

Conjugate: Unconjugated

Catalog #1: bs-51137R

Applications: WB, ELISA, IF, dot-blot

Reactivity: Human, Mouse

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

HeLa cells were stained with H3K27ac (1A3) Monoclonal Antibody (bsm-53022M) 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 H3K27ac antibody (left) diluted 1:500 in blocking solution followed by an anti-mouse antibody conjugated to Alexa594. The middle panel shows staining of the nuclei with DAPI. A merge of the two staining is shown on the right.
To test the specificity an ELISA was performed using a serial dilution of H3K27ac (1A3) Monoclonal Antibody (bsm-53022M). The wells were coated with peptides containing the unmodified H3K27 region as well as the acetylated H3K27 and the acetylated H3K9. Figure shows a high specificity of the antibody for the peptide containing the modification of interest.
ChIP assays were performed using HeLa cells, H3K27ac (1A3) Monoclonal Antibody (bsm-53022M) and optimized PCR primer pairs for qPCR. ChIP was performed using sheared chromatin from 1 million cells. A titration consisting of 0.5, 1, 2 and 5 \u00b5g of antibody per ChIP experiment was analyzed. IgG (1\u00b5g\/IP) was used as a negative IP control. Quantitative PCR was performed with primers specific for the promoters of the EIF4A2 and GAPDH genes, used as positive controls, and for 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).

Catalog #1: bsm-53022M

Applications: WB, ELISA, IF(ICC)

Reactivity: Human

Conjugate: Unconjugated

Catalog #1: bs-51021R

Applications: WB, ELISA, IF, dot-blot

Reactivity: Human

Conjugate: Unconjugated

Catalog #1: bs-51142R

Applications: WB, ELISA, IF, dot-blot

Reactivity: Human

Conjugate: Unconjugated

Catalog #1: bs-53142R

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

Catalog #1: bs-51108R

Applications: WB, ELISA, IF, dot-blot

Reactivity: Human

Conjugate: Unconjugated

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

Catalog #1: bs-53109R

Applications: WB, ELISA, IF, dot-blot

Reactivity: Human

Conjugate: Unconjugated

Catalog #1: bs-51109R

Applications: WB, ELISA, IF, dot-blot

Reactivity: Human

Conjugate: Unconjugated

Histone extracts (15 \u03bcg) from HeLa cells were analysed by Western blot using H3K27me3 (9B7) Monoclonal Antibody (bsm-53019M) 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 H3K27me3 (9B7) Monoclonal Antibody (bsm-53019M) and 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 H3K27me3 antibody (left) diluted 1:500 in blocking solution followed by an anti-mouse secondary 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 of H3K27me3 (9B7) Monoclonal Antibody (bsm-53019M). The wells were coated with peptides containing the unmodified H3K27 region as well as the mono-, di- and trimethylated H3K27 and the trimethylated H3K9. Figure shows a high specificity of the antibody for the peptide containing the modification of interest.
ChIP assays were performed using HeLa cells, H3K27me3 (9B7) Monoclonal Antibody (bsm-53019M) and optimized PCR primer sets for qPCR. ChIP was performed on sheared chromatin from 1 million cells. 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 promoters of the active genes c-fos and GAPDH as negative controls, and for the coding regions of the inactive genes MYT1 and TSH2B 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). These results are in accordance with the observation that H3K27me3 is preferably present at inactive genes.

Catalog #1: bsm-53019M

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

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

Catalog #1: bs-51019R

Applications: WB, ELISA, IF

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 the Bioss antibody against H3K27me3 (Cat. No. bs-53122R) diluted 1:500 in TBS-Tween containing 5% skimmed milk.
A Dot Blot analysis was performed to test the cross-reactivity of the Bioss antibody against H3K27me3 (Cat. No. bs-53122R) with peptides containing other modifications of histone H3 and H4 and the unmodified H3K27 sequence. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:5,000. Figure 4 shows a high specificity of the antibody for the modification of interest. Please note that that antibody also recognizes the modification if S28 is phosphorylated.
Mouse NIH3T3 cells were stained with the Bioss antibody against H3K27me3 (Cat. No. bs-53122R) and with DAPI. Cells were fixed with 4% formaldehyde for 10\u2019 and blocked with PBS\/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labeled with the H3K27me3 antibody (left) diluted 1:200 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.
To determine the titer of the antibody, an ELISA was performed using a serial dilution of H3K27me3 Polyclonal Antibody (Cat. No. bs-53122R). The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:3,500.

Catalog #1: bs-53122R

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

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

Conjugate: Unconjugated

Catalog #1: bs-51122R

Applications: WB, ELISA, IF, dot-blot

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

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

Catalog #1: bs-51053R

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

Reactivity: Human

Conjugate: Unconjugated

Catalog #1: bs-51132R

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

Catalog #1: bs-51131R

Applications: WB, ELISA, IF, dot-blot

Reactivity: Human

Conjugate: Unconjugated

To test the specificity an ELISA was performed using a serial dilution using 3K36me2 (6C9) Monoclonal Antibody (bsm-53020M). The wells were coated with peptides containing the unmodified H3K36 region as well as the mono-, di- and trimethylated H3K36 and the trimethylated H3K9. Figure shows a high specificity of the antibody for the peptide containing the modification of interest.
ChIP assays were performed using HeLa cells, H3K36me2 (6C9) Monoclonal Antibody (bsm-53020M) and optimized PCR primer pairs for qPCR. ChIP was performed using sheared chromatin from 1 million cells. A titration consisting of 0.5, 1, 2 and 5 \u03bcg of antibody per ChIP experiment was analyzed. IgG (1 \u03bcg\/IP) was used as a negative IP control. Quantitative PCR was performed with primers specific for a genomic region upstream of the TGM2 gene, used as a positive control, and for 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).

Catalog #1: bsm-53020M

Applications: ELISA, IF(ICC)

Reactivity: Human

Conjugate: Unconjugated

Catalog #1: bs-51020R

Applications: WB, ELISA, dot-blot

Reactivity: Human

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

Catalog #1: bs-51071R

Applications: WB, ELISA, IF, dot-blot

Reactivity: Human

Conjugate: Unconjugated

HeLa cells were stained with H3K36me3 (11D1) Monoclonal Antibody (bsm-53021M) and DAPI. Cells were fixed with methanol and blocked with PBS\/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labelled with the H3K36me3 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 of H3K36me3 (11D1) Monoclonal Antibody (bsm-53021M). The wells were coated with peptides containing unmodified H3K36 region as well as the mono-, di- and trimethylated H3K36 and the trimethylated H3K9. Figure shows a high specificity of the antibody for the peptide containing the modification of interest.
ChIP assays were performed using HeLa cells, H3K36me3 (11D1) Monoclonal Antibody (bsm-53021M) and optimized PCR primer pairs for qPCR. ChIP was performed using sheared chromatin from 1 million cells. A titration consisting of 0.5, 1, 2 and 5 \u03bcg of antibody per ChIP experiment was analyzed. IgG (1 \u03bcg\/IP) was used as a negative IP control. Quantitative PCR was performed with primers specific for the coding regions of the active EIF2S3 and CCT5 genes, used as positive controls, and for the inactive MYOD1 gene and the Sat2 satellite repeat, used as negative controls. 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-53021M

Applications: WB, ELISA, IF

Reactivity: Human

Conjugate: Unconjugated

A Dot Blot analysis was performed to test the cross-reactivity of the Bioss antibody against H3K36me3 (bs-53116R) with peptides containing other H3 and H4 modifications and the unmodified sequence. One hundred to 0.2 pmol of the peptide containing the respective histone modification were spotted on a membrane. The antibody was used at a dilution of 1:20,000. Figure shows a high specificity of the antibody for the modification of interest.
To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Bioss antibody directed against H3K36me3 (Cat. No. bs-53116R) and the crude serum. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution, the titer of the purified antibody was estimated to be 1:19,300.
ChIP was performed with 2 \u03bcg of the Bioss antibody against H3K36me3 (bs-53116R) on sheared chromatin from 1 million HeLaS3 cells using a ChIP-seq kit. IgG (2 \u03bcg\/IP) was used as a negative IP control. The IP\u2019d DNA was analyzed by qPCR with optimized PCR primer pairs for the promoter and coding region of the active GAPDH, for a region located 1 kb upstream of the GAPDH promoter and for the coding region of the active ACTB gene (figure A). The IP\u2019d DNA was subsequently analyzed on an Illumina Genome Analyzer. Library preparation, cluster generation, and sequencing were performed according to the manufacturer\u2019s instructions. The 36 bp tags were aligned to the human genome using the ELAND algorithm. Figure B shows the obtained profiles in genomic regions of chromosome 12 (including the GAPDH positive control), 7 (including the ACTB positive control), 14 and 3, respectively. These results clearly show an enrichment of the H3K36me3 at active genes.
ChIP assays were performed using HeLa cells, the Bioss antibody against H3K36me3 (Cat. No. bs-53116R) and optimized PCR primer sets for qPCR. ChIP was performed using sheared chromatin from 1 million cells on the SX-8G IP-Star automated system. A titration consisting of 1, 2, 5 and 10 \u03bcg of antibody per ChIP experiment was analyzed. IgG (2 \u03bcg\/IP) was used as a negative IP control. QPCR was performed with primers for the promoter and coding region of the active GAPDH, for a region located 1 kb upstream of the GAPDH promoter and for the Sat2 satellite repeat. The figure shows the recovery expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).

Catalog #1: bs-53116R

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

Reactivity: Human, Others, Zebrafish, Drosophila

Conjugate: Unconjugated