H4K8ac Polyclonal Antibody

A Dot Blot analysis was performed to test the cross-reactivity of the Bioss antibody against H4K8ac (cat. No. bs-53133R) with peptides containing other histone modifications and the unmodified H4K8. 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.

$Histone extracts of HeLa cells (15 \u03bcg) were analyzed by Western blot using the Bioss antibody directed against H4K8ac (cat. No. bs-53133R) diluted 1:200 in TBS-Tween containing 5% skimmed milk.$

Histone extracts of HeLa cells (15 \u03bcg) were analyzed by Western blot using the Bioss antibody directed against H4K8ac (cat. No. bs-53133R) diluted 1:200 in TBS-Tween containing 5% skimmed milk.

To determine the titer of the antibody, an ELISA was performed using a serial dilution of the Bioss antibody directed against H4K8ac (cat. No. bs-53133R), 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 1:16,700.

$Mouse NIH3T3 cells were stained with H4K8ac Polyclonal Antibody (cat. No. bs-53133R) and with DAPI. Cells were fixed with 4% formaldehyde for 10\u2019 and blocked with PBS\/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labeled with the H4K8ac antibody (left) diluted 1:500 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.$

Mouse NIH3T3 cells were stained with H4K8ac Polyclonal Antibody (cat. No. bs-53133R) 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 H4K8ac 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.

$ChIP assays were performed using human HeLa cells, H4K8ac Polyclonal Antibody (cat. No. bs-53133R) and optimized PCR primer sets for qPCR. ChIP was performed with a ChIP kit on sheared chromatin from 10,000 cells using the SX-8G IP-Star automated system. A titration of the antibody consisting of 1, 2, 5, and 10 \u03bcg per ChIP experiment was analyzed. IgG (1 \u03bcg\/IP) was used as negative IP control. QPCR was performed with primers for the c-fos promoter and for the Sat2 satellite repeat region. The figure shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).$

ChIP assays were performed using human HeLa cells, H4K8ac Polyclonal Antibody (cat. No. bs-53133R) and optimized PCR primer sets for qPCR. ChIP was performed with a ChIP kit on sheared chromatin from 10,000 cells using the SX-8G IP-Star automated system. A titration of the antibody consisting of 1, 2, 5, and 10 \u03bcg per ChIP experiment was analyzed. IgG (1 \u03bcg\/IP) was used as negative IP control. QPCR was performed with primers for the c-fos promoter and for the Sat2 satellite repeat region. 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
Mouse

Overview
Catalog #	bs-53133R
Product Name	H4K8ac Polyclonal Antibody
Applications	WB, ELISA, IF, dot-blot
Reactivity	Human, Mouse
Specifications
Conjugation	Unconjugated
Host	Rabbit
Source	Polyclonal antibody raised in rabbit against the region of histone H4 containing the acetylated lysine 8 (H4K8ac), using a KLH-conjugated synthetic peptide
Clonality	Polyclonal
Isotype	Not Determined
Concentration	1.2ug/ul
Purification	Affinity purified polyclonal antibody in PBS containing 0.05% azide and 0.05% ProClin 300.
Storage Buffer	PBS with 0.05% sodium azide and 0.05% Proclin300.
Storage Condition	Store at -20°C for 12 months.
Target
Gene ID	121504
Swiss Prot	P62805
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