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Formalin-fixed and paraffin embedded rat brain labeled with Rabbit Anti Histone H3 Polyclonal Antibody, Unconjugated (bs-0349R) at 1:200 followed by conjugation to the secondary antibody and DAB staining
Formalin-fixed and paraffin embedded mouse intestine labeled with Rabbit Anti Histone H3 Polyclonal Antibody, Unconjugated (bs-0349R) at 1:200 followed by conjugation to the secondary antibody and DAB staining
Formalin-fixed and paraffin-embedded rat brain labeled with Anti-Histone H3 Polyclonal Antibody, Unconjugated(bs-0349R) 1:200, overnight at 4\u00b0C, The secondary antibody was Goat Anti-Rabbit IgG, Cy3 conjugated(bs-0295G-Cy3)used at 1:200 dilution for 40 minutes at 37\u00b0C.
Mouse splenocytes probed with Rabbit Anti-Histone H3 Polyclonal Antibody, Unconjugated (bs-0349R) (green) at 1:50 for 30 minutes followed by a FITC conjugated secondary antibody compared to unstained cells (blue), secondary only(light blue), and isotype control(orange).

Catalog #1: bs-0349R

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

Reactivity: Human, Mouse, Rat, Others

Conjugate: Unconjugated

Formalin-fixed and paraffin embedded rat liver labeled with Rabbit Anti-MGMT Polyclonal Antibody, Unconjugated (bs-1002R) at 1:200 followed by conjugation to the secondary antibody and DAB staining
Formalin-fixed and paraffin embedded human lung cancer labeled with Rabbit Anti-MGMT Polyclonal Antibody, Unconjugated (bs-1002R) at 1:200 followed by conjugation to the secondary antibody and DAB staining

Catalog #1: bs-1002R

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

Reactivity: Rat, Others

Conjugate: Unconjugated

Western blot was performed on whole cell (25 \u00b5g, lane 1) and histone extracts (15 \u00b5g, lane 2) from HeLa cells, and on 1 \u00b5g of recombinant histone H2A, H2B, H3 and H4 (lane 3, 4, 5 and 6, respectively) using the Bioss antibody against H3K9me3 (Cat. No. bs-53114R). The antibody was diluted 1:1,000 in TBS-Tween containing 5% skimmed milk.
HeLa cells were stained with the Bioss antibody against H3K9me3 (bs-53114R) 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 H3K9me3 antibody (middle) diluted 1:1,000 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The left panel shows staining of the nuclei with DAPI. A merge of both stainings is shown on the right.
To determine the titer of the antibody, an ELISA was performed using a serial dilution of the antibody directed against human H3K9me3 (bs-53114R) in antigen-coated wells. The antigen used was a peptide containing the histone modification of interest. By plotting the absorbance against the antibody dilution, the titer of the antibody was estimated to be 1:30,000.
A Dot Blot analysis was performed to test the cross-reactivity of the antibody against H3K9me3 (Cat. No. bs-53114R) 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.

Catalog #1: bs-53114R

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

Reactivity: Human, Mouse, Others, Zebrafish

Conjugate: Unconjugated

Catalog #1: bs-0349R-PE

Applications: WB, FCM

Reactivity: Human, Mouse, Rat, Others

Conjugate: PE

Catalog #1: bs-0349R-FITC

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

Reactivity: Human, Mouse, Rat, Others

Conjugate: FITC

Catalog #1: bs-0349R-Cy5

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

Reactivity: Human, Mouse, Rat, Others

Conjugate: Cy5

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

Catalog #1: bs-53092R

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

Reactivity: Human, Mouse, Others

Conjugate: Unconjugated

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

Catalog #1: bs-53093R

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

Reactivity: Human, Mouse, Others

Conjugate: Unconjugated

Catalog #1: bs-0349R-Cy3

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

Reactivity: Human, Mouse, Rat, Others

Conjugate: Cy3

Catalog #1: bs-0349R-Cy7

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

Reactivity: Human, Mouse, Rat, Others

Conjugate: Cy7

Catalog #1: bs-0349R-Biotin

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

Reactivity: Human, Mouse, Rat, Others

Conjugate: Biotin

Catalog #1: bs-0349R-HRP

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

Reactivity: Human, Mouse, Rat, Others

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 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

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-0349r-cy55-100ul

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

Reactivity: Human, Mouse, Rat, Others

Conjugate: Cy5.5