Acetyl-Histone H4 (Lys12) Monoclonal Antibody

$4% Paraformaldehyde-fixed HeLa ( treated with Trichostatin A (TSA)(1\u03bcM, 18h) ) (H) cell; Triton X-100 at r.t. for 20 min; Antibody incubation with (Acetyl-Histone H4 (Lys12)) monoclonal Antibody, unconjugated (bsm-60055M) 1:100, 90 min at 37\u00b0C; followed by BF488 conjugated Goat Anti-Mouse IgG antibody (green, bs-60296G-BF488) at 37\u00b0C for 90 min, DAPI (blue, C02-04002) was used to stain the cell nuclei. PBS instead of the primary antibody was used as the blank control.$

4% Paraformaldehyde-fixed HeLa ( treated with Trichostatin A (TSA)(1\u03bcM, 18h) ) (H) cell; Triton X-100 at r.t. for 20 min; Antibody incubation with (Acetyl-Histone H4 (Lys12)) monoclonal Antibody, unconjugated (bsm-60055M) 1:100, 90 min at 37\u00b0C; followed by BF488 conjugated Goat Anti-Mouse IgG antibody (green, bs-60296G-BF488) at 37\u00b0C for 90 min, DAPI (blue, C02-04002) was used to stain the cell nuclei. PBS instead of the primary antibody was used as the blank control.

$Paraformaldehyde-fixed, paraffin embedded Rat Liver; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Acetyl-Histone H4 (Lys12) Monoclonal Antibody, Unconjugated (bsm-60055M) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-40296G-HRP and DAB (C-0010) staining.$

Paraformaldehyde-fixed, paraffin embedded Rat Liver; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Acetyl-Histone H4 (Lys12) Monoclonal Antibody, Unconjugated (bsm-60055M) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-40296G-HRP and DAB (C-0010) staining.

$Paraformaldehyde-fixed, paraffin embedded Human Liver; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Acetyl-Histone H4 (Lys12) Monoclonal Antibody, Unconjugated (bsm-60055M) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-40296G-HRP and DAB (C-0010) staining.$

Paraformaldehyde-fixed, paraffin embedded Human Liver; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Acetyl-Histone H4 (Lys12) Monoclonal Antibody, Unconjugated (bsm-60055M) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-40296G-HRP and DAB (C-0010) staining.

$Paraformaldehyde-fixed, paraffin embedded Rat Testicles; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Acetyl-Histone H4 (Lys12) Monoclonal Antibody, Unconjugated (bsm-60055M) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-40296G-HRP and DAB (C-0010) staining.$

Paraformaldehyde-fixed, paraffin embedded Rat Testicles; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Acetyl-Histone H4 (Lys12) Monoclonal Antibody, Unconjugated (bsm-60055M) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-40296G-HRP and DAB (C-0010) staining.

$Paraformaldehyde-fixed, paraffin embedded Human Testicles; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Acetyl-Histone H4 (Lys12) Monoclonal Antibody, Unconjugated (bsm-60055M) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-40296G-HRP and DAB (C-0010) staining.$

Paraformaldehyde-fixed, paraffin embedded Human Testicles; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Acetyl-Histone H4 (Lys12) Monoclonal Antibody, Unconjugated (bsm-60055M) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-40296G-HRP and DAB (C-0010) staining.

$Paraformaldehyde-fixed, paraffin embedded Rat Colon; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Acetyl-Histone H4 (Lys12) Monoclonal Antibody, Unconjugated (bsm-60055M) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-40296G-HRP and DAB (C-0010) staining.$

Paraformaldehyde-fixed, paraffin embedded Rat Colon; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Acetyl-Histone H4 (Lys12) Monoclonal Antibody, Unconjugated (bsm-60055M) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-40296G-HRP and DAB (C-0010) staining.

$Paraformaldehyde-fixed, paraffin embedded Rat Cerebrum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Acetyl-Histone H4 (Lys12) Monoclonal Antibody, Unconjugated (bsm-60055M) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-40296G-HRP and DAB (C-0010) staining.$

Paraformaldehyde-fixed, paraffin embedded Rat Cerebrum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Acetyl-Histone H4 (Lys12) Monoclonal Antibody, Unconjugated (bsm-60055M) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-40296G-HRP and DAB (C-0010) staining.

$Paraformaldehyde-fixed, paraffin embedded Human Cerebrum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Acetyl-Histone H4 (Lys12) Monoclonal Antibody, Unconjugated (bsm-60055M) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-40296G-HRP and DAB (C-0010) staining.$

Paraformaldehyde-fixed, paraffin embedded Human Cerebrum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Acetyl-Histone H4 (Lys12) Monoclonal Antibody, Unconjugated (bsm-60055M) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-40296G-HRP and DAB (C-0010) staining.

$Paraformaldehyde-fixed, paraffin embedded Rat Kidney; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Acetyl-Histone H4 (Lys12) Monoclonal Antibody, Unconjugated (bsm-60055M) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-40296G-HRP and DAB (C-0010) staining.$

Paraformaldehyde-fixed, paraffin embedded Rat Kidney; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Acetyl-Histone H4 (Lys12) Monoclonal Antibody, Unconjugated (bsm-60055M) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-40296G-HRP and DAB (C-0010) staining.

$Paraformaldehyde-fixed, paraffin embedded Human Kidney; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Acetyl-Histone H4 (Lys12) Monoclonal Antibody, Unconjugated (bsm-60055M) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-40296G-HRP and DAB (C-0010) staining.$

Paraformaldehyde-fixed, paraffin embedded Human Kidney; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Acetyl-Histone H4 (Lys12) Monoclonal Antibody, Unconjugated (bsm-60055M) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-40296G-HRP and DAB (C-0010) staining.

$Paraformaldehyde-fixed, paraffin embedded Rat Stomach; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Acetyl-Histone H4 (Lys12) Monoclonal Antibody, Unconjugated (bsm-60055M) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-40296G-HRP and DAB (C-0010) staining.$

Paraformaldehyde-fixed, paraffin embedded Rat Stomach; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Acetyl-Histone H4 (Lys12) Monoclonal Antibody, Unconjugated (bsm-60055M) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-40296G-HRP and DAB (C-0010) staining.

$Paraformaldehyde-fixed, paraffin embedded Human Stomach; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Acetyl-Histone H4 (Lys12) Monoclonal Antibody, Unconjugated (bsm-60055M) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-40296G-HRP and DAB (C-0010) staining.$

Paraformaldehyde-fixed, paraffin embedded Human Stomach; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Acetyl-Histone H4 (Lys12) Monoclonal Antibody, Unconjugated (bsm-60055M) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-40296G-HRP and DAB (C-0010) staining.

$Paraformaldehyde-fixed, paraffin embedded Rat Pancreas; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Acetyl-Histone H4 (Lys12) Monoclonal Antibody, Unconjugated (bsm-60055M) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-40296G-HRP and DAB (C-0010) staining.$

Paraformaldehyde-fixed, paraffin embedded Rat Pancreas; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Acetyl-Histone H4 (Lys12) Monoclonal Antibody, Unconjugated (bsm-60055M) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-40296G-HRP and DAB (C-0010) staining.

$Paraformaldehyde-fixed, paraffin embedded Human Lung Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Acetyl-Histone H4 (Lys12) Monoclonal Antibody, Unconjugated (bsm-60055M) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-40296G-HRP and DAB (C-0010) staining.$

Paraformaldehyde-fixed, paraffin embedded Human Lung Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; The section was incubated with Acetyl-Histone H4 (Lys12) Monoclonal Antibody, Unconjugated (bsm-60055M) at 1:200 overnight at 4\u00b0C, followed by conjugation to the bs-40296G-HRP and DAB (C-0010) staining.

$C2C12 (M) treated with Trichostatin A (TSA) (1\u03bcM) for 18 h, 25 \u03bcg total protein per lane of cell lysates (see on figure) probed with Acetyl-Histone H4 (Lys12) monoclonal antibody, unconjugated (bsm-60055M) at 1:1000 dilution and 4\u00b0C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.$

C2C12 (M) treated with Trichostatin A (TSA) (1\u03bcM) for 18 h, 25 \u03bcg total protein per lane of cell lysates (see on figure) probed with Acetyl-Histone H4 (Lys12) monoclonal antibody, unconjugated (bsm-60055M) at 1:1000 dilution and 4\u00b0C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.

$Chromatin immunoprecipitation was performed using cross-linked chromatin from 5\u00d710\u2076 HeLa cells (treated with 5 mM sodium butyrate for 24 hours) per IP, with 5 \u03bcg of bsm-60055M antibody or 5 \u03bcg of normal mouse IgG(negative control bs-0296P) , and 20 \u03bcL of Protein A\/G MagBeads per IP. The immunoprecipitated DNA was quantified by real-time PCR using primers specific for the human GAPDH-CDS, RPL30-promoter and RPL30-exon 3 regions. The data are presented as enrichment of each sample relative to the total amount of input at each amplicon.$

Chromatin immunoprecipitation was performed using cross-linked chromatin from 5\u00d710\u2076 HeLa cells (treated with 5 mM sodium butyrate for 24 hours) per IP, with 5 \u03bcg of bsm-60055M antibody or 5 \u03bcg of normal mouse IgG(negative control bs-0296P) , and 20 \u03bcL of Protein A\/G MagBeads per IP. The immunoprecipitated DNA was quantified by real-time PCR using primers specific for the human GAPDH-CDS, RPL30-promoter and RPL30-exon 3 regions. The data are presented as enrichment of each sample relative to the total amount of input at each amplicon.

Applications

WB
IHC-F
IF
IHC-P
ICC/IF

Reactivity

Human
Mouse
Rat

Overview
Catalog #	bsm-60055m
Product Name	Acetyl-Histone H4 (Lys12) Monoclonal Antibody
Applications	WB IHC-F IF IHC-P ICC/IF
Reactivity	Human, Mouse, Rat
Specifications
Conjugation	Unconjugated
Host	Mouse
Clonality	Monoclonal
Clone #	A8B8
Isotype	IgG
Concentration	Lot Dependent
Purification	Protein G coupled with the antigen peptide containing acetylated H4 (Lys12).
Storage Buffer	0.01M TBS(pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol.
Storage Condition	Store at -20°C for one year. Avoid repeated freeze/thaw cycles.
Target
Swiss Prot	P62805
Application Dilution
WB
IHC-F
IF
IHC-P
ICC/IF