Analysis Note

Negative ControlSK-OV-3 cells or normal skin tissue

Positive ControlA431 cells or breast carcinoma tissue

Application

Frozen sections (1 g/ml; see application references)Immunoblotting (0.1-1 g/ml; see application references)Immunocytochemistry (1-2.5 g/ml; see application references)Immunoprecipitation (1 g/ml or use Cat. No. OP43A; see application references)Paraffin sections (1 g/ml, pepsin, heat or pressure cooker pre-treatment required; see application references)

General description

Recognizes the ~53 kDa wild-type and mutant p53 protein in A431 cells and breast carcinoma tissue.

Purified mouse monoclonal antibody generated by immunizing BALB/c mice with the specified immunogen and fusing splenocytes with X63Ag8.653 mouse myeloma cells (see application references). Recognizes the ~53 kDa wild-type and mutant forms of p53.

This Anti-p53 (Ab-6) (Pantropic) Mouse mAb (DO-1) is validated for use in Frozen sections, Immunoblotting, ICC, IP, Paraffin sections for the detection of p53 (Ab-6) (Pantropic).

Immunogen

wild type, recombinant, human p53

Human

Epitope: Within amino acids 21-25 of human p53

Legal Information

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

Other Notes

Recognizes both mutant and wild-type p53 under denaturing and non-denaturing conditions. This antibody reacts weakly with rodent p53; we do not recommend it for rodent samples. For gel shift assay, use Cat. No. OP43L resuspended in 100 µl of buffer. Wild-type p53 has a short half life and is present in low amounts in cells. For immunoprecipitation increasing the amount of sample and labeling with ³⁵S-Met for less than or equal to 1 h will aid in visualizing wild-type p53. 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.Kastan, M.B., et al. 1992. Cell71, 587.Kuerbitz, S.J. 1992. Proc. Natl. Acad. Sci. USA89, 7491.Lane, D.P. 1992. Nature358, 15.Vojtesek, B., et al. 1992. J. Immunol. Meth.151, 237.Kastan, M.B., et al. 1991 Cancer Res.51, 6304.

Packaging

Please refer to vial label for lot-specific concentration.

20, 100 µg in Plastic ampoule

Physical form

In 0.05 M sodium phosphate buffer, 0.2% gelatin.

Warning

Toxicity: Standard Handling (A)