Analysis Note

Positive ControlA431 or SVT2 cells or breast carcinoma tissue

Application

Frozen Sections (1:500)Immunoblotting (1:2500)Immunocytochemistry (1:500)Paraffin Sections (1:500, pepsin or heat pre-treatment required)Immunoprecipitation (chromatin, see application references)

Components

Anti-p53 antisera, normal sheep serum, and biotinylated rabbit anti-sheep IgG

General description

Recognizes the ~53 kDa wild-type and mutant p53 protein in A431 and SVT2 cells and breast carcinoma tissue.

Sheep polyclonal antibody supplied as diluted serum. Recognizes the ~53 kDa wild-type or mutant p53 protein. Supplied with normal sheep serum and biotinylated rabbit anti-sheep IgG.

This Anti-p53 (Ab-7) (Pantropic) Sheep pAb is validated for use in Frozen Sections, Immunoblotting, ICC, Paraffin Sections, IP for the detection of p53 (Ab-7) (Pantropic).

Immunogen

recombinant, human p53 protein

Human

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. USA 89, 7491.Lane, D.P. 1992. Nature358, 15.Kastan, M.B., et al. 1991. Cancer Res.51, 6304.

Recommended dilutions for the biotinylated rabbit anti-sheep secondary are 1:25,000-1:200,000 for immunoblotting and 1:10,000 for immunocytochemistry and paraffin sections. For immunoblotting, do not add normal serum, non-fat dried milk, culture medium or other potential sources of biotin to the working solutions (i.e. block with 2-5% fatty acid free BSA). Use streptavidin-horseradish peroxidase (Cat. No. OR03L) or streptavidin-alkaline phosphatase (Cat. No. OR04L) at 10 ng/ml to attach an enzyme to the biotinylated secondary antibody, then visualize using standard colorimetric or chemiluminescent reagents. To maximize sensitivity of immunoblots, preconcentrate samples by immunoprecipitation with Cat. No. OP03, OP09, or OP43, then immunoblot using Cat. No. PC35 and chemiluminescent detection. Antibodies should be titrated for optimal results in individual systems.

Packaging

1 ea in Plastic ampoule

Physical form

200 µl diluted (1:4) anti-p53 sheep serum, 500 µl undiluted normal sheep serum, and 100 µl biotinylated rabbit anti-sheep IgG.

Warning

Toxicity: Standard Handling (A)