ChIPAb+ p53 - ChIP Validated Antibody and Primer Set

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+ p53 set includes the p53 antibody raised against recombinant human p53, the negative control antibody (mouse normal IgG), and qPCR primers flanking an Sp1 binding site (to which p53 can be recruited1) within the human p21 (WAF1/CIP1/CDKN1A) promoter, amplifying a 105 base pair PCR product. The p53 and negative control antibodies are supplied in a scalable "per ChIP" reaction size and can be used to functionally validate the precipitation of p53 associated chromatin.

Specificity

Recognizes p53 in wild type and mutant forms of human origin.

Application

Chromatin Immunoprecipitation Analysis:
Sonicated chromatin prepared from untreated or actinomycin D treated (30 ng/mL, 6 hrs.) U2OS cells (3 X 10⁶ cell equivalents per IP) was subjected to chromatin immunoprecipitation using 1 μg of mouse Anti-p53 and the Magna ChIP^® G (Cat. # 17-611) Kit. Immunoprecipitation of p53 associated DNA fragments was verified by qPCR using ChIP Primers p21 flanking the human p21 promoter that contains an Sp1 binding site (Please see figures).
Fold Increase is a ratio of normalized mean IP quantities extracted from standard curves derived from inputs of each chromatin sample. As previously published, p53 association with this region of the p21 promoter increases upon actinomycin D treatment.

Chromatin Immunoprecipitation Analysis:
Sonicated chromatin prepared from untreated or UV treated (6 hrs, 50 joules/m2.) U2OS cells (3 X 10⁶ cell equivalents per IP) was subjected to chromatin immunoprecipitation using 1 μg of mouse Anti-p53 and the Magna ChIP A (Cat. # 17-610) Kit.
Immunoprecipitation of p53 associated DNA fragments was verified by qPCR using ChIP Primers p21 flanking the human p21 promoter that contains an Sp1 binding site (Please see figures).
Fold Increase is a ratio of normalized mean IP quantities extracted from standard curves derived from inputs of each chromatin sample. p53 immunoprecipitable activity associated with this promoter increases with UV treatment as observed in other studies.

Research Category
Epigenetics & Nuclear Function

Research Sub Category
Chromatin Biology

This ChIPAb+ p53 -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers.

Packaging

25 assays per kit. ~1 μg per chromatin immunoprecipitation

Quality

Routinely evaluated by chromatin immunoprecipitation and immunoblot on RIPA lysate of human Raji cells.

Target description

53 kDa

Physical form

Anti-p53 (mouse monoclonal IgG). One vial containing 25 μg of of protein A purified mouse IgG2a in 25 μL of storage buffer (PBS, pH 7.4, containing 0.05% sodium azide and 1 mg/mL BSA). Store at -20°C.

Normal Mouse IgG. One vial containing 25 μg of normal mouse IgG in 25 μL volume. Store at -20°C.

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. Store at -20°C.
FOR: GTG GCT CTG ATT GGC TTT CTG
REV: CTG AAA ACA GGC AGC CCA AG

Format: Purified

Protein A purified

Storage and Stability

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

Analysis Note

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

Legal Information

MAGNA CHIP is a registered trademark of Merck KGaA, Darmstadt, Germany

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.