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 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.

Specificity

Acetyl-Histone H3 (Lys9)

Immunogen

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

Application

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.

Packaging

25 assays per kit, ~5μg per chromatin immunoprecipitation

Quality

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.

Target description

17 kDa

Physical form

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

Storage and Stability

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

Analysis Note

Control
Included negative control antibody purified rabbit IgG and control primers specific for human p21 (WAF1/CIP1/CDKN1A) promoter.

Legal Information

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

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.

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