1. Since applications vary, each investigator should titrate the reagent to obtain optimal results. 2. Please refer to us for technical protocols. 3. 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.
Purification
The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.
Applications include western blot analysis (1-2 µg/ml), immunoprecipitation (1-2 µg/1 x 10^6 cells), immunofluorescence microscopy of cultured cells, immunohistochemistry of frozen (5-20 µg/ml), and antigen-unmasked paraffin-embedded tissue sections (5-20 µg/ml). Positive control cell lines include SK-BR-3 human breast carcinoma cells (ATCC HTB-30), and A431 human vulval carcinoma cells (ATCC CRL-1555). COS-7 SV40 transformed monkey kidney cells (ATCC CRL-1651) or another SV40-transformed cell line are also useful as positive controls for detecting p53. MCF-7 human breast carcinoma cells (ATCC HTB-22) are suggested as a negative control. Positive immunostaining is seen in a high proportion of breast and colon carcinomas. p53 staining is not typically detected in normal skin, brain, kidney, lung, stomach or breast tissue.
This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Storage
4 °C
Storage Comment
Store undiluted at 4°C.
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The gene for the nuclear phosphoprotein 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 over-expressed in a variety of transformed cells and it forms specific complexes with several viral oncogenes including SV40 large T, E1B from adenovirus and E6 from human papilloma virus. Recent data suggest that wild type p53 plays a role as a checkpoint protein for DNA damage during the 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. p53 migrates at a reduced molecular weight of 53 kDa. Clone PAb 1801 recognizes an epitope between amino acids 32-79 in the N-terminal domain of human wild type and mutant p53 antibody. It does not cross-react with p53 from other species. A truncated recombinant human p53 fusion protein was used as immunogen.