Description General description Purified mouse monoclonal antibody generated by immunizing BALB/c mice with the specified immunogen and fusing splenocytes with SP2 mouse myeloma cells (see application references). Recognizes the ~53 kDa mutant p53 protein under non-denaturing conditions. Recognizes both the mutant and the wild-type p53 protein under denaturing conditions. Recognizes the ~53 kDa mutant p53 protein under non-denaturing conditions by immunoprecipitation, immunofluorescence, and flow cytometry. Recognizes both mutant and wild-type p53 by immunoblotting and paraffin sections under denaturing conditions. This Anti-p53 (Ab-3) (Mutant) Mouse mAb (PAb240) is validated for use in FC, Frozen Sections, Gel Shift, Immunoblotting, IF, IP, Paraffin Sections for the detection of p53 (Ab-3) (Mutant). Immunogen Epitope: within amino acids 213-217 Human a recombinant protein consisting of amino acids 14-389 of p53 fused to β-galactosidase Application Flow Cytometry (1-20 µg/ml) Frozen Sections (10 µg/ml) Gel Shift (see comments) Immunoblotting (5 µg/ml) Immunofluorescence (1-20 µg/ml, see application references) Immunoprecipitation (1 µg per sample) Paraffin Sections (see application references) Physical form In 50 mM sodium phosphate buffer, 0.2% gelatin, pH 7.5. Packaging 100, 200 μg in Plastic ampoule Please refer to vial label for lot-specific concentration. Warning Toxicity: Standard Handling (A) Analysis Note Negative Control SK-OV-3 cells Positive Control A431, Hs27 (wild-type p53), or SK-BR-3 cells or breast carcinoma tissue Other Notes El-Deiry, W.S., et al. 1994. Cancer Res.54, 1169. Greenblatt, M.S., et al. 1994. Cancer Res.54, 4855. Barak, Y., et al. 1993. EMBO J.12, 461. Kastan, M.B., et al. 1992. Cell71, 587. Kuerbitz, S.J. 1992. Proc. Natl. Acad. Sci. USA89, 7491. Lane, D.P. 1992. Nature358, 15. Kastan, M.B., et al. 1991. Cancer Res.51, 6304. Under non-denaturing conditions (immunoprecipitation, immunofluorescence and frozen sections), Anti-p53 (Ab-3) does not recognize normal (wild-type) p53 protein; it recognizes an epitope exposed by activating mutations or denaturation. In denaturing protocols (immunoblotting and paraffin sections), Anti-p53 (Ab-3) will recognize both mutant and wild-type p53. Will not recognize to some p53 molecules with mutations in the RHSVV epitope, but will react to TFIIIA, which has the RHSVV epitope. For gel shift assay, use Cat. No. OP29L and resuspend in 100 µl buffer. Antibody should be titrated for optimal results in individual systems.