PE Mouse Anti-p53 Antibody Set

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BD Pharmingen™ PE Mouse Anti-p53 Antibody Set

Name: PE Mouse Anti-p53 Antibody Set
Brand: BD Pharmingen™
SKU: 556534

(RUO)

Protocol Library

Scientific Resources

Profile of permeabilized HT-29 colon adenocarcinoma cells analyzed on a FACScan™ (BDIS, San Jose, CA). Cells were stained with anti-human p53-PE (clone DO-7) or with an IgG2b isotype control.

Protocol Library

Scientific Resources

Product Details

Brand: BD Pharmingen™

Reactivity: Human (QC Testing), Monkey, Cow (Tested in Development)

Application: Intracellular staining (flow cytometry) (Routinely Tested)

Regulatory Status: RUO

RRID: AB_396454

Description

p53 is a 53 kDa nuclear phosphoprotein that acts as a tumor suppressor protein, and is involved in inhibiting cell proliferation when DNA damage occurs. The gene for p53 is the most commonly mutated gene yet identified in human cancers. Missense mutations occur in tumors of the colon, lung, breast, ovary, bladder and several other organs. The mutant p53 is overexpressed in a variety of transformed cells and wild-type p53 forms specific complexes with several viral oncogenes including SV40 large T, E1B from adenovirus, and E6 from human papilloma virus. Wild type p53 plays a role as a checkpoint protein for DNA damage during the G1/S-phase of the cell cycle. However, it is still unclear, whether point mutated forms of p53 are simple null mutants and/or dominant negatively acting proteins.

Clone DO-7 recognizes human wildtype and mutant p53. It cross-reacts with bovine p53 but does not cross-react with mouse or rat p53. Clone DO-7 recognizes an epitope between amino acids 1-45 of all known forms of human p53. Human recombinant p53 protein was used as immunogen. Wildtype 53 proteins have a very short half-life and are usually not detectable with monoclonal antibodies in normal tissues. Clone DO-7 was initially characterized by immunoprecipitation, western blot analysis, immunohistochemistry of frozen and formalin-fixed paraffin-embedded tissue sections, and flow cytometric analysis.

Clone 27-35 is an isotype control monoclonal antibody specific for hapten dansyl (5-[dimethylamino] naphthalene-1-sulfonyl). This hapten is not expressed on human cells or human cell lines.

Preparation And Storage

Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with R-PE under optimum conditions, and unconjugated antibody and free PE were removed.

Recommended Assay Procedures

Positive control cell lines include SK-BR-3 human breast carcinoma cells (ATCC HTB-30), HT-29 human colon adenocarcinoma (ATCC HTB-38) and A431 human vulval carcinoma cells (ATCC CRL-1555). MCF-7 human breast carcinoma cells (ATCC HTB-22) are suggested as a negative control.

Product Notices

This antibody has been optimized and preassayed with its matched isotype control to be used at the recommended volume of 20 ul/test. Titration of the reagents or substituting with other (non-matched) isotype control is NOT recommended.
Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
Source of all serum proteins is from USDA inspected abattoirs located in the United States.
Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.

556534 Rev. 10

Components

Description	Quantity/Size	Part Number	EntrezGene ID
PE Mouse Anti-Human p53	100 Tests (1 ea)	51-15805X	N/A
PE Mouse IgG2b, κ Isotype Control	100 Tests (1 ea)	51-66375X-2	N/A

556534 Rev. 10

Citations & References

Development References (7)

Baas IO, Mulder JW, Offerhaus GJ, Vogelstein B, Hamilton SR. An evaluation of six antibodies for immunohistochemistry of mutant p53 gene product in archival colorectal neoplasms. J Pathol. 1994; 172(1):5-12. (Clone-specific: Immunohistochemistry). View Reference
Bonsing BA, Corver WE, Gorsira MC, et al. Specificity of seven monoclonal antibodies against p53 evaluated with Western blotting, immunohistochemistry, confocal laser scanning microscopy, and flow cytometry. Cytometry. 1997; 28(1):11-24. (Clone-specific: Flow cytometry). View Reference
Cripps KJ, Purdie CA, Carder PJ, et al. A study of stabilisation of p53 protein versus point mutation in colorectal carcinoma. Oncogene. 1994; 9(9):2739-2743. (Clone-specific: Immunohistochemistry). View Reference
Jacquemier J, Moles JP, Penault-Llorca F, et al. p53 immunohistochemical analysis in breast cancer with four monoclonal antibodies: comparison of staining and PCR-SSCP results. Br J Cancer. 1994; 69(5):846-852. (Clone-specific: Immunohistochemistry). View Reference
Lambkin HA, Mothersill CM, Kelehan P. Variations in immunohistochemical detection of p53 protein overexpression in cervical carcinomas with different antibodies and methods of detection. J Pathol. 1994; 172(1):13-18. (Clone-specific: Immunohistochemistry). View Reference
Vogelstein B. Cancer. A deadly inheritance. Nature. 1990; 348(6303):681-682. (Biology). View Reference
Vojtesek B, Bartek J, Midgley CA, Lane DP. An immunochemical analysis of the human nuclear phosphoprotein p53. New monoclonal antibodies and epitope mapping using recombinant p53. J Immunol Methods. 1992; 151(1-2):237-244. (Clone-specific: Immunohistochemistry, Immunoprecipitation, Western blot). View Reference

View All (7)

556534 Rev. 10

Please refer to Support Documents for Quality Certificates

Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described

Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.

For Research Use Only. Not for use in diagnostic or therapeutic procedures.

Refer to manufacturer's instructions for use and related User Manuals and Technical Data Sheets before using this product as described.

Comparisons, where applicable, are made against older BD technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.