Merck, OP29L-100UG, Anti-p53 (Ab-3) (Mutant) Mouse mAb (PAb240)

General description
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.
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.
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).
Immunogen
Epitope: within amino acids 213-217
Human
a recombinant protein consisting of amino acids 14-389 of p53 fused to β-galactosidase
Application
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)
Physical form
Lyophilized from a volatile buffer, 100 µg BSA.
Preparation Note
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
Analysis Note
Negative Control
SK-OV-3 cells
Positive Control
A431, Hs27 (wild-type p53), or SK-BR-3 cells or breast carcinoma
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. USA89, 7491.
Lane, D.P. 1992. Nature358, 15.
Kastan, M.B., et al. 1991. Cancer Res.51, 6304.
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.
Legal Information
CALBIOCHEM is a registered trademark of Merck KGaA, Darmstadt, Germany
Disclaimer
Toxicity: Standard Handling (A)