H3K9acS10p Polyclonal Antibody

$Histone extracts of HeLa cells (15 \u03bcg) were analysed by Western blot using the H3K9acS10p antibody (bs-53062R) diluted 1:250 in TBS-Tween containing 5% skimmed milk. The position of the protein of interest is indicated on the left; the marker (in kDa) is shown on the right.$

Histone extracts of HeLa cells (15 \u03bcg) were analysed by Western blot using the H3K9acS10p antibody (bs-53062R) 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.

A Dot Blot analysis was performed to test the cross reactivity of the H3K9acS10p antibody (bs-53062) with peptides containing other modifications of histone H4 and H3 or unmodified histone H3 sequences. 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 double modification.

Immunofluorescence using the H3K9acS10p antibody at 1:500 dilution in Hela cells.

$ChIP assays were performed using human osteosarcoma (U2OS) cells, the H3K9acS10p antibody (bs-53062R) and optimized PCR primer sets for qPCR. ChIP was performed using sheared chromatin from 1.5 million cells. A titration of the antibody consisting of 5 and 15 \u03bcl per ChIP experiment was analysed. IgG (5 \u03bcg\/IP) was used as negative IP control. QPCR was performed with primers for the ALDOA (fructose-bisphosphate aldolase A) promoter and for the coding region of the myogenic differentiation gene (MYOD), a gene that is inactive at normal conditions. The figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).$

ChIP assays were performed using human osteosarcoma (U2OS) cells, the H3K9acS10p antibody (bs-53062R) and optimized PCR primer sets for qPCR. ChIP was performed using sheared chromatin from 1.5 million cells. A titration of the antibody consisting of 5 and 15 \u03bcl per ChIP experiment was analysed. IgG (5 \u03bcg\/IP) was used as negative IP control. QPCR was performed with primers for the ALDOA (fructose-bisphosphate aldolase A) promoter and for the coding region of the myogenic differentiation gene (MYOD), a gene that is inactive at 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).

Applications

WB
ELISA
IF
dot-blot

Reactivity

Human

Overview
Catalog #	bs-53062R
Product Name	H3K9acS10p Polyclonal Antibody
Applications	WB, ELISA, IF, dot-blot
Reactivity	Human
Specifications
Conjugation	Unconjugated
Host	Rabbit
Source	Polyclonal antibody raised in rabbit against histone H3 containing the acetylated lysine 9 and the phosphorylated serine 10 (H3K9acS10p), using a KLH-conjugated synthetic peptide
Clonality	Polyclonal
Isotype	Not Determined
Concentration	Not Determined
Purification	Whole antiserum from rabbit containing 0.05% azide
Storage Buffer	Serum with 0.05% sodium azide.
Storage Condition	Store at -20°C for 12 months.
Target
Gene ID	8350
Swiss Prot	P68431
Background	Core component of nucleosome. Nucleosomes wrap and compact DNA into chromatin, limiting DNA accessibility to the cellular machineries which require DNA as a template. Histones thereby play a central role in transcription regulation, DNA repair, DNA replication and chromosomal stability. DNA accessibility is regulated via a complex set of post-translational modifications of histones, also called histone code, and nucleosome remodeling.
Application Dilution
WB	1:300-5000
ELISA	1:500-1000
IF
dot-blot