Analysis Note

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

Negative ControlNormal skin tissue or SK-OV-3 cells

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

Purified mouse monoclonal antibody, originally called clone L21, generated by immunizing mice with the specified immunogen and fusing splenocytes with NS1 mouse myeloma cells (see application references). Recognizes the ~53 kDa wild-type and mutant forms of p53.

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

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

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.

Originally called clone L21. Does not recognize phosphorylated or acetylated p53. For a gel shift assay, 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 wild-type p53, maximize sensitivity by concentrating samples by immunoprecipitation with Cat. No. OP03, then perform immunoblotting using Cat. No. PC35 and chemiluminescent detection. Antibody should be titrated for optimal results in individual systems.

Packaging

100 µg in Plastic ampoule

Physical form

Lyophilized from a volatile buffer, 100 µg BSA.

Reconstitution

Reconstitute the lyophilized antibody with sterile PBS, pH 7.4, or sterile 20 mM Tris-saline (20 mM Tris containing 0.15 M NaCl), pH 7.4, to yield a final concentration of 100 µg/ml. Lyophilized antibody should be resuspended at 4°C with occasional gentle mixing for at least 2 h. Store at 4°C until reconstituted, then store in aliquots at -20°C or at 4°C with 0.1% azide; freezing of aliquots is best for long-term storage of reconstituted product, but repetitive freezing and thawing should be avoided.

Warning

Toxicity: Standard Handling (A)