H3K27ac (1A3) Monoclonal Antibody

$HeLa cells were stained with H3K27ac (1A3) Monoclonal Antibody (bsm-53022M) and with DAPI. Cells were fixed with 4% formaldehyde for 10\u2019 and blocked with PBS\/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labelled with the H3K27ac antibody (left) diluted 1:500 in blocking solution followed by an anti-mouse antibody conjugated to Alexa594. The middle panel shows staining of the nuclei with DAPI. A merge of the two staining is shown on the right.$

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.

$ChIP assays were performed using HeLa cells, H3K27ac (1A3) Monoclonal Antibody (bsm-53022M) and optimized PCR primer pairs for qPCR. ChIP was performed using sheared chromatin from 1 million cells. A titration consisting of 0.5, 1, 2 and 5 \u00b5g of antibody per ChIP experiment was analyzed. IgG (1\u00b5g\/IP) was used as a negative IP control. Quantitative PCR was performed with primers specific for the promoters of the EIF4A2 and GAPDH genes, used as positive controls, and for the inactive MYOD1 gene and the Sat2 satellite repeat, used as negative controls. Figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).$

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).

Applications

WB
ELISA
IF(ICC)

Reactivity

Human

Overview
Catalog #	bsm-53022M
Product Name	H3K27ac (1A3) Monoclonal Antibody
Applications	WB, ELISA, IF(ICC)
Reactivity	Human
Specifications
Conjugation	Unconjugated
Host	Mouse
Source	Monoclonal antibody raised in mouse against histone H3 acetylated at lysine 27 (H3K27ac), using a KLHconjugated synthetic peptide.
Clonality	Monoclonal
Clone #	1A3
Isotype	IgG1
Concentration	1ug/ul
Purification	Protein A purified monoclonal antibody in PBS containing 0.05% azide.
Storage Buffer	PBS with 0.05% sodium azide.
Storage Condition	Store at -20°C for 12 months.
Target
Background	Histones are the main constituents of the protein part of chromosomes of eukaryotic cells. They are rich in the amino acids arginine and lysine and have been greatly conserved during evolution. Histones pack the DNA into tight masses of chromatin. Two core histones of each class H2A, H2B, H3 and H4 assemble and are wrapped by 146 base pairs of DNA to form one octameric nucleosome. Histone tails undergo numerous post-translational modifications, which either directly or indirectly alter chromatin structure to facilitate transcriptional activation or repression or other nuclear processes. In addition to the genetic code, combinations of the different histone modifications reveal the so-called histone code. Histone methylation and demethylation is dynamically regulated by respectively histone methyl transferases and histone demethylases.
Application Dilution
WB	1:300-5000
ELISA	1:500-1000
IF(ICC)	1:50-200