ELISA, Immunoprecipitation, Western Blot. 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 p21WAF1 and not p21 complexes with other proteins. High level expression of p21WAF1 requires induction of wild-type p53 expression (e.g. via exposure to DNA damaging agents).
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Description
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.
Immunogen
This protein A purified monoclonal antibody was produced by repeated immunizations with recombinant human p21 fusion protein.
Yeh, C. H. and Shatkin, A.J. (1995). A cis-acting element in Rous sarcoma virus long terminal repeat required for promoter repression by Hela nuclear protein p21. J. Biol. Chem. 270:15815-15820 Joshi, U. S. et al. (1998). Inhibition of tumor cell growth by p21WAF1 adenoviral gene transfer in lung cancer. Cancer Gene Ther. 5: 183-191.Yang, Z.Y. et al. (1995) The p21 cyclin-dependent kinase inhibitor suppresses tumorigenicity in vivo. Nat Med 1(10):1052-1056Kagawa, S. et al. (1997) p53 expression overcomes p21WAF1/CIP1-mediated G1 arrest and induces apoptosis in human cancer cells. Oncogene 15(16):1903-1909.Waldman, et al. (1995) p21 is necessary for the p53-mediated G1 arrest in human cancer cells. Cancer Res 55(22):5187-5190.Kovalic, J. et al., (1996) Int.J.Oncol. 9, suppl., 835.Stephane Supiot, Richard P. Hill and Robert G. Bristow (2008) Nutlin-3 radiosensitizes hypoxic prostate cancer cells independent of p53. Mol Cancer Ther 7; 993.