Anti-p53 (Ab-6) (Pantropic) Mouse mAb (DO-1)

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 mutant and wild-type forms of p53.

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

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

Epitope: Within amino acids 21-25 of human p53

Human

wild type, recombinant, human p53

Frozen sections (1 µg/ml; see application references)

Gel Shift (see comments)

Immunoblotting (0.1 µg/ml; see application references)

Immunocytochemistry (1-2.5 µg/ml; see application references)

Immunoprecipitation (1 µg/ml or use Cat. No. OP43A; see application references)

Paraffin sections (1 µg/ml, pepsin, heat or pressure cooker pre-treatment required; see application references)

Lyophilized from a volatile buffer, 100 µg BSA.

We recommend resuspending the lyophilized antibody with sterile phosphate buffered saline (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. Reconstituted product will be more stable if 0.1% sodium azide is added (do not add azide if antibody is to be used with viable cells). Lyophilized antibodies should be resuspended at 4°C with occasional gentle mixing for at least two hours. Following reconstitution, refrigerate (4°C) for short-term storage or aliquot and freeze (-20°C) for long-term storage.

Negative Control
SK-OV-3 cells or normal skin tissue

Positive Control
A431 cells or breast carcinoma tissue

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.

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, resuspend 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.

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

Toxicity: Standard Handling (A)