| Clone: | BP53-12 |
|---|---|
| Isotype: | Mouse IgG2a |
| Specificity: | The antibody BP53-12 recognizes defined epitope (aa 16-25) on human p53, a 50 kDa tumour suppressor found in increased amounts in a wide variety of transformed cells; it is frequently mutated or inactivated in many types of cancer. |
| Immunogen: | Bacterially expressed full-length wild-type p53 |
| Species Reactivity: | Human, Non-Human Primates |
| Application: |
Immunoprecipitation
Western Blotting
Recommended dilution: 1-2 μg/ml, overnight in 4°C
Positive control: RAMOS human lymphoma cell line Sample preparation: Resuspend approx. 50 mil. cells in 1 ml cold Lysis buffer (1% laurylmaltoside in 20 mM Tris/Cl, 100 mM NaCl pH 8.2, 50 mM NaF including Protease inhibitor Cocktail). Incubate 60 min on ice. Centrifuge to remove cell debris. Mix lysate with non-reducing SDS-PAGE sample buffer. Application note: Non-reducing conditions. SDS-PAGE (12% separating gel).
Immunohistochemistry (paraffin sections)
Immunocytochemistry
ELISA
|
| Purity: | > 95% (by SDS-PAGE) |
| Purification: | Purified from ascites by precipitation methods. |
| Concentration: | 1 mg/ml |
| Storage Buffer: | Phosphate buffered saline (PBS) with 15 mM sodium azide, approx. pH 7.4 |
| Storage / Stability: | Store at 2-8°C. Do not use after expiration date stamped on vial label. For long-term storage aliquot and store at -20°C. Avoid freeze/thaw cycles. |
| Expiration: | See vial label |
| Lot Number: | See vial label |
| Background: | The tumour suppressor protein p53 is a key element of intracellular anticancer protection. It mediates cell cycle arrest or apoptosis in response to DNA damage or to starvation for pyrimidine nukleotides. It is up-regulated in response to these stress signals and stimulated to activate transcription of specific genes, resulting in expression of p21waf1 and other proteins involved in G1 or G2/M arrest, or proteins that trigger apoptosis, such as Bcl-2. The structure of p53 comprises N-terminal transactivation domain, central DNA-binding domain, oligomerisation domain, and C-terminal regulatory domain. There are various phosphorylation sites on p53, of which the phosphorylation at Ser15 is important for p53 activation and stabilization. |
| References: |
*Agarwal ML, Agarwal A, Taylor WR, Stark GR: p53 controls both the G2/M and the G1 cell cycle checkpoints and mediates reversible growth arrest in human fibroblasts. Proc Natl Acad Sci U S A. 1995 Aug 29;92(18):8493-7.
*Agarwal ML, Agarwal A, Taylor WR, Chernova O, Sharma Y, Stark GR: A p53-dependent S-phase checkpoint helps to protect cells from DNA damage in
response to starvation for pyrimidine nucleotides. Proc Natl Acad Sci U S A. 1998 Dec 8;95(25):14775-80.
*Taylor WR, DePrimo SE, Agarwal A, Agarwal ML, Schönthal AH, Katula KS, Stark GR: Mechanisms of G2 arrest in response to overexpression of p53. Mol Biol Cell. 1999 Nov;10(11):3607-22.
*Taylor WR, Agarwal ML, Agarwal A, Stacey DW, Stark GR: p53 inhibits entry into mitosis when DNA synthesis is blocked. Oncogene. 1999 Jan 14;18(2):283-95.
*Tanigawa S, Fujii M, Hou DX: Stabilization of p53 is involved in quercetin-induced cell cycle arrest and
apoptosis in HepG2 cells. Biosci Biotechnol Biochem. 2008 Mar;72(3):797-804.
*Bartek J, Bartkova J, Vojtesek B, Staskova Z, Lukas J, Rejthar A, Kovarik J, Midgley CA, Gannon JV, Lane DP: Aberrant expression of the p53 oncoprotein is a common feature of a wide spectrum of human malignancies. Oncogene. 1991 Sep;6(9):1699-703.
*Bartkova J, Bartek J, Lukas J, Vojtesek B, Staskova Z, Rejthar A, Kovarik J, Midgley CA, Lane DP: p53 protein alterations in human testicular cancer including pre-invasive intratubular germ-cell neoplasia. Int J Cancer. 1991 Sep 9;49(2):196-202.
*Dolezalova H, Vojtesek B, Kovarik J: Epitope analysis of the human p53 tumour suppressor protein. Folia Biol (Praha). 1997;43(1):49-51.
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