Purification and Quality Control The His tagged C-terminus-deleted p53 (amino acid 1-363) was expressed in a baculovirus system and purified by affinity and FPLC chromatography.
Unit Definition (Activity) 1 unit equals 1 nanogram of purified protein. Variation exist in different lots.
Applications Recombinant p53 can be used for: 1) gel mobility shift assay or for a DNase I footprinting in the presence of double stranded DNA containing a consensus p53-binding sequence [5’-PuPuPuC(A/T)(T/A)GPyPyPy-3’]; 2) in vitro transcription assay; 3) protein-protein interaction assay; and 4) cell growth assay.
Formulation and Storage This protein was kept in 20 mM Tris-Cl (pH 8.0), 20% glycerol, 100 mM KCl, 1 mM DTT and 0.2 mM EDTA. Always store at -80˚C, avoid repeated freeze thaw cycles.
Synonym Homo sapiens Mdm2 p53 binding protein homolog (mouse) (MDM2); hdm2; HDMX; MGC5370 and MGC71221.
Background Human p53 protein is composed of 393 amino acid residues with several distinct regions. The N-terminal activation domain allows p53 protein to recruit the basal transcription machinery and activate the expression of target genes. The core domain binds to target DNA in a sequence-specific manner and the majority of mutations found in human tumors occur in the region of the gene encoding this domain (1-3). The C-terminal domain is composed of predominantly basic residues and modification of the C-terminal basic domain, including acetylation, glycosylation and phosphorylation, is an essential mechanism for regulating p53 function (4-6). Disruption or loss of oligomerization function is associated with loss of cell cycle arrest (5, 6). This mutant protein (with the deletion of the C-terminus 51 residues including the entire basic domain and a portion of the tetramerization domain) can be used as a unique tool to study specific functions of p53 related to the C-terminus.
References: 1. Pellegata, NS. et al., (1995) Oncogene 11, 337-349 2. El-Deiry, WS. et al., (1992) Nature Genet 1, 45-49 3. Hollstein, M. et al., (1991) Science 253, 49-53 4. Hupp, TR. et al., (1992) Cell 71, 875-886 5. Ishioka, C. et al., (1995) Oncogene 10, 1485-1492 6. Waterman, MJ. et al., (1996) Cancer Res 56, 158-163
DISCLAIMER
This products is recommended For RESEARCH USE ONLY and is Not qualified for Use in Diagnostic or Therapeutic Procedures.
