Analysis Note

Positive ControlHs27 cells or breast carcinoma tissue

Application

Frozen Sections (see applications references)r>Immunoblotting (not recommended)r>Immunofluorescence (1-20 µg/ml)r>Immunoprecipitation (1 µg per sample)r>Paraffin Sections (5 µg/ml, heat pre-treatment required)

General description

This Anti-p53 (Ab-5) (Wild type) Mouse mAb (PAb1620) is validated for use in Frozen Sections, Immunoblotting, IF, IP, Paraffin Sections for the detection of p53 (Ab-5) (Wild type).

Purified mouse monoclonal antibody generated by immunizing BALB/c mice with the specified immunogen and fusing splenocytes with SP2/0 Ag14 myeloma cells. Recognizes the ~53 kDa wild-type p53 protein.

Recognizes the ~53 kDa wild-type p53 protein in its native conformation in Hs27 cells and breast carcinoma tissue. Does not recognize mutant or denatured p53 protein.

Immunogen

vLM mouse tumor cells

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.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.Ball, R.K., et al. 1984. EMBO J.3, 1485.

Wild-type p53 has a short half-life and is present in low amounts in cells. Increasing the amount of sample to be immunoprecipitated and applied to the gel may help for visualization. Short incubation times with ³⁵S-Met(≤ 1 hr) will help reduce background. p53 (Ab-5) may also be used to visualize p53 in cytospins or cultured cells using standard horseradish peroxidase or immunofluorescence detection techniques. p53 (Ab-5) preferentially immunoprecipitates wild-type p53; it should not precipitate mutant or denatured p53. Antibody should be titrated for optimal results in individual systems.

Packaging

Please refer to vial label for lot-specific concentration.

20, 100 µg in Plastic ampoule

Warning

Toxicity: Standard Handling (A)