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Key Specifications Table
| Species Reactivity | Key Applications |
|---|---|
| H | WB, ChIP |
| Description | |
|---|---|
| Catalogue Number | 17-613 |
| Brand Family | Upstate |
| Trade Name |
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| Description | ChIPAb+ p53 - ChIP Validated Antibody and Primer Set |
| Overview | 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. |
| Product Information | |
|---|---|
| Format | Purified |
| Control |
|
| Presentation | 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 |
| Quality Level | MQ100 |
| Applications | |
|---|---|
| Application | This ChIPAb+ p53 -ChIP Validated Antibody & Primer Set conveniently includes the antibody & the specific control PCR primers. |
| Key Applications |
|
| Application Notes | 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. |
| Biological Information | |
|---|---|
| Host | Mouse |
| Specificity | Recognizes p53 in wild type and mutant forms of human origin. |
| Isotype | IgG |
| Species Reactivity |
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| Antibody Type | Monoclonal Antibody |
| Entrez Gene Number |
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| Gene Symbol |
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| Purification Method | Protein A Purfied |
| UniProt Number |
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| Molecular Weight | 53 kDa |
| Product Usage Statements | |
|---|---|
| Quality Assurance | Routinely evaluated by chromatin immunoprecipitation and immunoblot on RIPA lysate of human Raji cells. |
| Usage Statement |
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| Storage and Shipping Information | |
|---|---|
| Storage Conditions | Stable for 1 year at -20°C from date of receipt |
| Packaging Information | |
|---|---|
| Material Size | 25 assays |
| Material Package | 25 assays per kit. ~1 μg per chromatin immunoprecipitation |