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About This Item

UNSPSC Code:
12352203
NACRES:
NA.43
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biological source

mouse

Quality Level

100

antibody form

purified antibody

antibody product type

primary antibodies

clone

PAb240, monoclonal

form

lyophilized

does not contain

preservative

species reactivity

hamster, rat, mouse, bovine, chicken, human

should not react with

Xenopus

manufacturer/tradename

Calbiochem®

storage condition

OK to freeze

dilution

(Flow Cytometry (1-20 µg/mL)
Frozen Sections (10 µg/mL)
Gel Shift
Immunoblotting (5 µg/mL)
Immunofluorescence (1-20 µg/mL)
Immunoprecipitation (1 µg per sample)
Paraffin Sections )

isotype

IgG1

shipped in

ambient

storage temp.

2-8°C

target post-translational modification

unmodified

Gene Information

human ... TP53(7157)

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)

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Storage Class Code

11 - Combustible Solids

WGK

WGK 1

Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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