Analysis Note

Positive ControlBreast carcinoma, A431 cells, or p53 standard (Cat. No. 506147)

Negative ControlSK-OV-3 cells or normal skin

Application

Frozen Sections (10 µg/ml, see application references)r>Gel Shift (see comments)r>Immunoblotting (10 µg/ml, see application references)r>Immunofluorescence (1-20 µg/ml, see application references)r>Flow Cytometry (1-20 µg/ml)r>Immunoprecipitation (1 µg per sample, see application references)

General description

Recognizes the ~53 kDa mammalian wild-type and mutant p53 protein. Does not recognize phosphorylated or acetylated p53.

Purified mouse monoclonal antibody generated by immunizing mice with the specified immunogen and fusing splenocytes with NS1 mouse myeloma cells (see application references). Recognizes the ~53 kDa mutant and wild-type p53 proteins.

This Anti-p53 (Ab-1) (Pantropic) Mouse mAb (PAb421) is validated for use in Frozen Sections, Gel Shift, Immunoblotting, IF, FC, IP for the detection of p53 (Ab-1) (Pantropic).

Immunogen

partially purified mouse p53

Epitope: within amino acids 376-378 of human p53

Mouse

Legal Information

CALBIOCHEM is a registered trademark of Merck KGaA, Darmstadt, Germany

Other Notes

Originally called clone L21. Does not recognize phosphorylated p53. For a gel shift assay, use Cat. No. OP03L and resuspend in 100 µl buffer. Wild-type p53 has a short half-life and is present in low amounts in cells. For immunoprecipitation, increasing the amount of sample to be immunoprecipitated and applied to the gel may help visualize wild-type p53, and short incubation times with ³⁵S-Met (≤1 h) will help reduce background. For immunoblotting of wild-type p53, maximize sensitivity by concentrating samples by immunoprecipitation with Cat. No. OP03, then immunoblot using PC35 and chemiluminescent detection. This antibody recognizes the region encompassing Ser³⁷⁶ to Ser³⁷⁸ (see application references) and does not recognize phosphorylated p53 (see application references). Antibody should be titrated for optimal results in individual systems.

El-Deiry, W.S., et al. 1994. Cancer Res.54, 1169.Greenblatt, M.S., et al. 1994. Cancer Res.54, 4855.Legros, Y., et al. 1994. Oncogene9, 2071.Barak, Y., et al. 1993. EMBO J.12, 461.Kuerbitz, S.J. 1992. Proc. Natl. Acad. Sci. USA89, 7491.Kastan, M.B., et al. 1992. Cell71, 587.Lane, D.P. 1992. Nature358, 15.Kastan, M.B., et al. 1991. Cancer Res.51 6304.Crawford, L. and Harlow, E. 1982. J. Virol.41, 709.Harlow, E., et al. 1981. J. Virol.39 861.

Packaging

20, 100 µg in Glass bottle

Please refer to vial label for lot-specific concentration.

Physical form

In 50 mM sodium phosphate buffer, 0.2% gelatin, pH 7.5.

Warning

Toxicity: Standard Handling (A)