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OP29L
Sigma-AldrichAnti-p53 (Ab-3) (Mutant) Mouse mAb (PAb240)
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).
More>>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). Less<<
Anti-p53 (Ab-3) (Mutant) Mouse mAb (PAb240) MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.
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.
Catalogue Number
OP29L
Brand Family
Calbiochem®
References
References
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.
Product Information
Form
Lyophilized
Formulation
Lyophilized from a volatile buffer, 100 µg BSA.
Negative control
SK-OV-3 cells
Positive control
A431, Hs27 (wild-type p53), or SK-BR-3 cells or breast carcinoma
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)
Application Comments
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.
Biological Information
Immunogen
a recombinant protein consisting of amino acids 14-389 of p53 fused to β-galactosidase
Immunogen
Human
Epitope
within amino acids 213-217
Clone
PAb240
Host
Mouse
Isotype
IgG₁
Species Reactivity
Bovine
Chicken
Hamster
Human
Mouse
Rat
Antibody Type
Monoclonal Antibody
Storage and Shipping Information
Ship Code
Ambient Temperature Only
Toxicity
Standard Handling
Storage
+2°C to +8°C
Do not freeze
Ok to freeze
Special Instructions
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
Note that this data sheet is not lot-specific and is representative of the current specifications for this product. Please consult the vial label and the certificate of analysis for information on specific lots. Also note that shipping conditions may differ from storage conditions.
Revision
26-February-2008 JSW
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)
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.
Background
The human p53 tumor suppressor gene encodes a 393 amino acid phosphoprotein that exhibits sequence-specific DNA binding and directly interacts with various cellular and viral proteins. p53 is the most commonly mutated gene in human cancer, with the majority of the mutations being amino acid substitutions. The normal function of p53 is to effect cell cycle arrest at the G1 and G2 checkpoints in response to DNA damage. This checkpoint function is executed by accumulation of p53 followed by induction of the GADD45, WAF1 and MDM2 genes. The current model of p53 function postulates that p53 senses DNA damage and arrests the cell cycle in either the G1 or G2 phases to allow DNA repair to take place. If repair is not successful, p53 initiates programmed cell death, thus preventing the propagation of genetic defects to successive generations of cells.
Host
Mouse
Immunogen species
Human
Immunogen
a recombinant protein consisting of amino acids 14-389 of p53 fused to β-galactosidase
Epitope
within amino acids 213-217
Clone
PAb240
Isotype
IgG₁
Species
not Xenopus, bovine, chicken, hamster, human, mouse, rat
Positive control
A431, Hs27 (wild-type p53), or SK-BR-3 cells or breast carcinoma
Negative control
SK-OV-3 cells
Form
Lyophilized
Formulation
Lyophilized from a volatile buffer, 100 µg BSA.
Preservative
None
Comments
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.
Storage
+2°C to +8°C
Do Not Freeze
Ok to freeze
Special Instructions
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
Toxicity
Standard Handling
References
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.
Application references
Paraffin Sections
Pezzella, F., et al. 1994. J. Clin. Pathol.47, 592.
Original Clone
Gannon, J.V., et al. 1990. EMBO J.9, 1595.
Epitope Identification
Stephen, C.W. and Lane, D.P., 1992. J. Mol. Biol.225, 1.
Immunofluorescence
Bernardi, R., et al. 2004. Nat. Cell Biol.6, 665.
