Cyclin-Dependent Kinase Inhibitor 1A (p21, Cip1) (CDKN1A) antibody

Antigen: Cyclin-Dependent Kinase Inhibitor 1A (p21, Cip1) (CDKN1A)

Synonyms: P21, CIP1, SDI1, WAF1, CAP20, CDKN1, MDA-6, p21CIP1, CDKI, Waf1, mda6, Cdkn1, p21WAF, p21Cip1, Cip1, CDKN1A, p21, cip1, p16Xic2, sdi1, waf1, cap20, cdkn1, mda-6, p21cip1, si:ch1073-48m5.2

Clonality: Monoclonal (WA-1)

Host: Mouse

Reactivity: Human, Mouse (Murine), Rat (Rattus), Sheep (Ovine)

Application: ELISA, Western Blotting (WB), Immunoprecipitation (IP)

Catalog no.: ABIN105216

Additional Information

Alternative name: p21/WAF1

Immunogen: This protein A purified monoclonal antibody was produced by repeated immunizations with recombinant human p21 fusion protein. Hybridoma: Produced by the fusion between BALB/c mouse splenocytes and mouse myeloma SP2 cells using conventional hybridoma technology.

Format: Liquid (sterile filtered)

Isotype: IgG1 kappa  (Matching secondary antibodies)

Clone: WA-1

Description: P21 WAF1 is a tumor suppressor gene product critical to maintain proper regulatory control of cell division. p21 WAF1 inhibits cyclin/cyclin-dependent kinase (cdk) complexes, causing cell cycle arrest by blocking cell division at the S phase. Various mutations of tumor suppressor genes have been associated with different types of cancers. In these cases, somatic gene therapy with wild-type versions of tumor suppressor genes have been contemplated as anti-cancer therapeutic approaches. p21 WAF1 contains p53-binding sites in its promoter and expression of p21 WAF1 is induced in WI38 fibroblasts with PMA. The induction of and accumulation of p21 WAF1 does not affect p53 levels. BRCA1 limits tumor growth by inducing p21 WAF1 . Anti- p21 WAF1 has also referred to as anti-CIP1, anti-p21, anti-Sdi1, anti-Pic1, anti-CAP20 and anti-WAF.

Specificity: This protein A purified mouse monoclonal antibody reacts with human and mammalian p21. The intracellular protein p21 is a 21 kD protein and also known as wild-type p53-activated fragment 1 (WAF1). It is an inhibitor of cyclin-dependent kinases (Cdks) and proliferating-cell nuclear antigen (PCNA). The antibody is highly specific and shows no cross-reactivity with other closely related mitotic inhibitors.

Application Details

Application Notes: Recommended Dilutions: This product was assayed by immunoblot using HRP Goat-anti-Mouse IgG and ECL for detection. A working dilution of 1:500 to 1:1,000 is suggested for this assay. For immunohistochemistry dilute the antibody 1:50. Researchers should determine optimal titers for other applications. Note: This antibody is suitable for immunohistochemistry ELISA and immunoblotting. The antibody detects a 21 kDa band corresponding to the p21 protein. MCF7 cells or UV treated fibroblasts are suggested as a positive control for immunoblotting and immunohistochemistry. Methanol/acetone fixed tissue is reactive for immunohistochemistry. The antibody also reacts with breast carcinoma on frozen sections (methanol/acetone fixation) and on paraffin sections (formalin fixation and microwave treatment prior to staining). In ELISA this monoclonal antibody is useful either as a capture antibody (attached to the solid phase) or for detection of p21 protein. This antibody immunoprecipitates monomeric p21 WAF1 and not p21 complexes with other proteins. (This antibody co-precipitates all known p21 WAF1 interacting proteins including PCNA, cdk1, cdk2 and cdk4.) High level expression of p21 WAF1 requires induction of wild-type p53 expression (e.g. via exposure to DNA damaging agents).

Concentration: 1.0 mg/ml (by UV absorbance at 280 nm)

Buffer: Stabilizer: None. Preservative: 0.01% (w/v) Sodium Azide. Buffer: 0.02 M Potassium Phosphate, 0.5 M Sodium Chloride, pH 7.2.

Storage: Store vial at -20° C prior to opening. Aliquot contents and freeze at -20° C or below for extended storage. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4° C as an undiluted liquid. Dilute only prior to immediate use. Expiration date is one (1) year from date of opening.

Research Area: Cancer, Cell Cycle

Restrictions: For Research Use only

Images

Mab anti-Human p21WAF1antibody (clone WA-1) is shown to detect human p21 by western blot. Detection occurs after 10 ug of a HeLa whole cell lysate is loaded per lane. The blot was incubated with a 1:1,000 dilution of Mab anti-Human p21WAF1at room temperature for 30 min followed by detection using IRDye

Publications

Publications: Supiot, Hill, Bristow: "Nutlin-3 radiosensitizes hypoxic prostate cancer cells independent of p53." in: Molecular cancer therapeutics, Vol. 7, Issue 4, pp. 993-9, 2008 (PubMed).