Mouse anti P53 (TP53 tumor suppressor protein)

P53 is a tumor suppressor gene expressed in a wide variety of tissue types and is involved in regulating cell growth, replication, and apoptosis. It binds to MDM2, SV40 T antigen and Human papilloma virus E6 protein. Alterations of the p53 tumor suppressor gene are considered critical events in multistage carcinogenesis of a wide range of Human cancers. The mutations found in sporadic cancer are commonly missense mutations that inactivate the tumor suppressor activity of p53. These mutant proteins are much more stable than the normal p53 protein, frequently adopt an altered conformation and accumulate within the nucleus of the malignant cells. Mutations involving p53 are found in a wide variety of malignant tumors, including breast, ovarian, bladder, colon, lung, and melanoma. Positive nuclear staining with p53 antibodies has been reported to be a negative prognostic factor in amongst others breast carcinoma, lung carcinoma, colorectal, and urothelial carcinoma.

DO-1 is a mouse monoclonal IgG2a antibody derived by fusion of mouse myeloma Sp2/0-Ag14 cells with spleen cells from BALB/c mice immunized with recombinant Human wild type p53 protein expressed in E. coli.

The DO-1 antibody is suitable for the detection of p53 by Western blotting and detects a band of 53kDa in tumor tissue and cell extracts._x000D_ The antibody is suitable for immunocytochemistry on permeabilized cells, and Immunohistochemistry on frozen and formalin fixed, paraffin embedded tissues. _x000D_ Staining of formalin-fixed tissues requires boiling tissue sections in 10mM citrate buffer, pH 6.0, for 10-20 min followed by cooling at RT for 20 minutes._x000D_ The antibody can also be applied in ELISA for the quantification of the p53 protein._x000D_ Optimal antibody dilutions for the different applications should be determined by titration. Recommended range is 1:500 – 1:1000 for flow cytometry, 1:500-1:2000 for immunohistochemistry, and 1:500 – 1:1000 for immunoblotting applications.

Each vial contains 100 µL 1 mg/mL purified monoclonal antibody in phosphate buffered saline (PBS) containing 0,05% sodium azide DO-1 recognizes a 53kDa protein, which is identified as p53 suppressor gene product; It reacts with the mutant as well as the wild form of p53; Its epitope maps within the N-terminus (amino acids 11-25) of p53

_x000D_ Caution and warning:_x000D_ This product is intended FOR RESEARCH USE ONLY, and FOR TESTS IN VITRO, not for use in diagnostic or therapeutic procedures involving Humans or animals. This product contains sodium azide. To prevent formation of toxic vapors, do not mix with strong acidic solutions. To prevent formation of potentially explosive metallic azides in metal plumbing, always wash into drain with copious quantities of water._x000D_ This datasheet is as accurate as reasonably achievable, but Nordic-MUbio accepts no liability for any inaccuracies or omissions in this information.

_x000D_ 1. Vojtĕsek B, Bártek J, Midgley CA, Lane DP._x000D_ An immunochemical analysis of the Human nuclear phosphoprotein p53. New monoclonal antibodies and epitope mapping using recombinant p53._x000D_ J Immunol Methods. 1992;151(1-2):237-44._x000D_ _x000D_ 2. Vojtĕsek B, Fisher CJ, Barnes DM, Lane DP._x000D_ Comparison between p53 staining in tissue sections and p53 proteins levels measured by an ELISA technique. Br J Cancer. 1993;67(6):1254-8._x000D_ _x000D_ 3. Stephen CW, Helminen P, Lane DP._x000D_ Characterisation of epitopes on Human p53 using phage-displayed peptide libraries: insights into antibody-peptide interactions._x000D_ J Mol Biol. 1995;248(1):58-78.

The antibody is shipped at ambient temperature and may be stored at +4°C; For prolonged storage prepare appropriate aliquots and store at or below -20°C; Prior to use, an aliquot is thawed slowly in the dark at ambient temperature, spun down again and used to prepare working dilutions by adding sterile phosphate buffered saline (PBS, pH 7.2); Repeated thawing and freezing should be avoided; Working dilutions should be stored at +4°C, not refrozen, and preferably used the same day; If a slight precipitation occurs upon storage, this should be removed by centrifugation; It will not affect the performance or the concentration of the product