Anti-p53 (Ab-3) (Mutant) Mouse mAb (PAb240)

100

mouse

purified antibody

primary antibodies

PAb240, monoclonal

lyophilized

preservative

mouse, hamster, human, rat, chicken, bovine

Calbiochem^®

OK to freeze

IgG1

ambient

2-8℃

100 μg in Plastic ampoule

Purified mouse monoclonal antibody generated by immunizing BALB/c mice with the specified immunogen and fusing of splenocytes with SP2 mouse myeloma cells (see application references). Recognizes the ~53 kDa mutant p53 protein under non-denaturing conditions and both mutant and wild-type p53 under denaturing conditions.
This Anti-p53 (Ab-3) (Mutant) Mouse mAb (PAb240) is validated for use in FC, Frozen Sections, Gel Shift, Immunoblotting, IF, IP, Paraffin Sections for the detection of p53 (Ab-3) (Mutant).
Recognizes the ~53 kDa mutant p53 protein under non-denaturing conditions by immunoprecipitation, immunofluorescence, and flow cytometry. Recognizes both mutant and wild-type p53 by immunoblotting and paraffin sections under denaturing conditions.

Human
Epitope: within amino acids 213-217
a recombinant protein consisting of amino acids 14-389 of p53 fused to β-galactosidase

Flow Cytometry (1-20 µg/ml)

Frozen Sections (10 µg/ml)

Gel Shift (see comments)

Immunoblotting (5 µg/ml)

Immunofluorescence (1-20 µg/ml)

Immunoprecipitation (1 µg per sample)

Paraffin Sections (see application references)

Toxicity: Standard Handling (A)

Lyophilized from a volatile buffer, 100 µg BSA.

Reconstitute the lyophilized antibody with sterile 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. Lyophilized antibodies should be reconstituted at 4°C with occasional gentle mixing for at least 2 h. Following reconstitution, refrigerate (4°C) for short-term storage or aliquot and freeze (-20°C) for long-term storage with 0.1% azide

Positive Control
A431, Hs27 (wild-type p53), or SK-BR-3 cells or breast carcinoma
Negative Control
SK-OV-3 cells

Under non-denaturing conditions (immunoprecipitation, immunofluorescence and frozen sections), p53 (Ab-3) does not bind to normal (wild-type) p53 protein; it recognizes an epitope exposed by activating mutations or denaturation. In denaturing protocols (immunoblotting and paraffin sections), p53 (Ab-3) will recognize both mutant and wild-type p53. Will not recognize to some p53 molecules with mutations in the RHSVV epitope, but will recognize TFIIIA, which has the RHSVV epitope. For gel shift assay, resuspend in 100 µl buffer. Antibodies 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.

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