Analysis Note

Positive ControlA431 cells or breast carcinoma tissue

Negative ControlSK-OV-3 cells or normal skin

Application

Frozen Sections (5 µg/ml)r>Gel Shift (see comments)r>Immunoblotting (2.5 µg/ml)r>Immunoprecipitation (1 µg/sample)r>Paraffin Sections (5 µg/ml, pepsin, trypsin, or heat pre-treatment required)

General description

This Anti-p53 (Ab-2) (Pantropic) Mouse mAb (PAb1801) is validated for use in Frozen Sections, Gel Shift, Immunoblotting, IP, Paraffin Sections for the detection of p53 (Ab-2) (Pantropic).

Recognizes the ~53 kDa wild-type and mutant p53 protein in A-431 cells and breast carcinoma tissue.

Purified mouse monoclonal antibody generated by immunizing BALB/c mice with the specified immunogen and fusing splenocytes with SP2/0 Ag14 mouse myeloma cells (see application references). Recognizes the ~53 kDa mutant and wild-type forms of p53.

Immunogen

a human p53 fusion protein

Epitope: within amino acids 46-55 of human p53

Human

Legal Information

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

Other Notes

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; short incubation times with ³⁵S-Met (≤ 1 h) will help reduce background. For immunoblots of wild-type p53, maximize sensitivity by preconcentrating samples by immunoprecipitation with Cat. No. OP09, then immunoblot using Cat. No. PC35 and chemiluminescent detection. For a gel shift assay, use Cat. No. OP09L and resuspend in 100 µl buffer. 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.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.

Packaging

100, 200 µg in Plastic ampoule

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)