Flow Cytometry Immunoprecipitation
Western Blotting
Recommended dilution: 1-2 µg/ml, overnight in 4oC
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: 2-10ug/ml
ELISA

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.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.