ChIPAb+ Acetyl-Histone H3 (Lys9) Purified - ChIP Validated Antibody and Primer Set

17 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 H3 (Lys9) set includes the anti-acetyl-histone H3 (Lys9) antibody, a negative control antibody (purified Rabbit IgG), and qPCR primers flanking an Sp1 binding site in the human p21 (WAF1/CIP1/CDKN1A) promoter, amplifying a 105 base pair PCR product. The acetyl-histone H3 (Lys9) and negative control antibodies are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of acetyl-histone H3 (Lys9) associated chromatin.

The acetyl-histone H3 (Lys9) purified antibody is made against a peptide (acetylated at Lys9) corresponding to amino acids 1-12 of histone H3.

Chromatin Immunoprecipitation:
Sonicated chromatin prepared from untreated or UV treated (50 J/m2, 6 hrs.) U2OS cells (3 X 10⁶ cell equivalents per IP) was subjected to chromatin immunoprecipitation using 5 μg of either a normal rabbit IgG or Anti-Acetyl-Histone H3 (Lys9) antibody and the Magna ChIP A (Cat. #17-610) Kit. Successful immunoprecipitation of acetyl-histone H3 (Lys9) associated DNA fragments was verified by qPCR using control ChIP Primers p21 flanking the human p21(WAF1/CIP1/CDKN1A) promoter (Please see figures). Data is presented as fold enrichment of normalized percent input of each IP sample relative to input treated or untreated chromatin.
Please refer to the EZ-Magna A ChIP (Cat. #17-408) or EZ-ChIP (Cat. #17-371) protocol for experimental details.

Western Blot Analysis:
Acid-extracted 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 PVDF membrane and probed with Anti-Acetyl Histone H3 (Lys9) (1 μg/mL). Proteins were visualized using a goat-anti rabbit secondary antibody conjugated to HRP and a chemiluminescence detection system (Please see figures).

Research Category
Epigenetics & Nuclear Function

Research Sub Category
Chromatin Biology

This ChIPAb+ Acetyl-Histone H3 (Lys9) Purified -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.

Acetyl-Histone H3 (Lys9)

25 assays per kit, ~5μg per chromatin immunoprecipitation

Anti-Acetyl-Histone H3 (Lys9) (rabbit polyclonal IgG). One vial containing 125 μg of protein A purified antibody in 125 μL storage buffer containing 0.05% sodium azide and 30% glycerol.

Normal Rabbit IgG. One vial containing 125 ug purified Rabbit IgG in 125 uL storage buffer containing 0.05% sodium azide.

ChIP Primers p21. One vial containing 75 μL of 5 μM of each primer specific for a region of the human p21 (WAF1/CIP1/CDKN1A) promoter.
FOR: GTG GCT CTG ATT GGC TTT CTG
REV: CTG AAA ACA GGC AGC CCA AG

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

Chromatin Immunoprecipitation:
Sonicated chromatin prepared from UV treated (50 J/m2, 6 hrs.) U2OS cells (3 X 10⁶ cell equivalents per IP) were subjected to chromatin immunoprecipitation using 5 μg of either a normal rabbit IgG or Anti-Acetyl-Histone H3 (Lys9) antibody and the Magna ChIP A (Cat. #17- 610) Kit. Successful immunoprecipitation of acetyl-histone H3 (Lys9) associated DNA fragments was verified by qPCR using control ChIP Primers p21 flanking the human p21(WAF1/CIP1/CDKN1A) 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 antibody purified rabbit IgG and control primers specific for human p21 (WAF1/CIP1/CDKN1A) promoter.

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