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Analysis Note

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

Application

Research Sub CategoryHistones

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

Research CategoryEpigenetics & Nuclear Function

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).

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

General description

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.

Immunogen

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

Legal Information

UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany

Packaging

25 assays per set. ~4 µL per chromatin immunoprecipitation

Physical form

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.

Quality

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.

Specificity

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.

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

Storage and Stability

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

Target description

10 kDa