ChIPAb+ Acetyl-Histone H4 - ChIP Validated Antibody and Primer Set

10 kDa

All ChIPAb+ antibodies are individually validated for chromatin precipitation, every lot, every time. Each ChIPAb+ antibody set includes control primers (tested every lot by qPCR) to biologically validate your IP results in a locus-specific context. The qPCR protocol and primer sequences are provided, allowing researchers to validate ChIP protocols when using our antibody in their chromatin context. Each set also includes a negative control antibody to ensure specificity of the ChIP reaction.
The ChIPAb+ Acetyl-Histone H4 set includes purified rabbit polyclonal antiserum and the normal rabbit IgG antiserum, which can be used to demonstrate that the acetyl-histone H4 antibody is capable of precipitating acetyl-histone H4 associated chromatin. The qPCR primers included flank the human GAPDH promoter and produce a 166 base pair PCR product.

The acetyl-histone H4 antiserum is made against a peptide corresponding to amino acids 2-19 of Tetrahymena histone H4.

Research Category
Epigenetics & Nuclear Function

Research Sub Category
Histones

This ChIPAb+ Acetyl-Histone H4 -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.

Western Blot Analysis:
Acidextracted proteins from normal HeLa cells (Lane 1) and HeLa cells treated with 5 mM sodium butyrate for 24 hours (Lane 2) were resolved by electrophoresis, transferred to nitrocellulose and probed with anti-acetyl Histone H4 (1:2000).
Proteins were visualized using a goat-anti rabbit secondary antibody conjugated to HRP and a chemi-luminescence detection system. (Please see figures).

Predicted broad cross-reactivity among eukaryotes based upon sequence similarity.

Recognizes acetylated histone H4 of approximately 10 kDa. Cross-reacts with acetylated histone H2B from Tetrahymena and weakly crossreacts with acetylated histone H2B from HeLa cells.
May cross-react with other acetylated proteins.

25 assays per set. ~4 μL per chromatin immunoprecipitation

Anti-Acetyl-Histone H4 (rabbit polyclonal IgG). One vial containing 100 μL of antiserum containing 0.05% sodium azide. Store at -20°C.

Normal Rabbit Serum. One vial containing 100 uL antiserum containing 0.05% sodium azide. Store at -20°C.

Control Primers. One vial containing 75 μL of 5 μM of each primer specific for for human GAPDH.

Stable for 1 year at -20°C from date of receipt.

Chromatin Immunoprecipitation:
Sonicated chromatin prepared from untreated HeLa cells (1 X 10⁶ cell equivalents) was subjected to chromatin immunoprecipitation using 4 μL of either a normal rabbit antiserum or Anti-Acetyl Histone
H4 serum and the Magna ChIP A (Cat. # 17-610) Kit. Successful immunoprecipitation of acetyl histone H4 associated DNA fragments was verified by qPCR using control ChIP Primers flanking the human GAPDH promoter (Please see figures).
Please refer to the EZ-Magna A ChIP (Cat. # 17-408) or EZ-ChIP (Cat. # 17-371) protocol for experimental details.

Control
Included negative control rabbit IgG antiserum and control primers specific for human GAPDH promoter.

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