GST-tagged N terminal p53(1-81, Cat# p4002-01, 10ug)
mutated N terminal p53(1-81, L22A, W23A, Cat# p4003-01, 10ug)
p53 gene access number is NM_000546
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, whereas 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).
The His tagged wild type and C-terminus-deleted p53 (amino acid 1-363 and amino acid 1-343) were expressed and purified from the baculovirus systems. GST-tagged N terminal wild type and mutated N termianl domain of p53(1-81) were expressed and purified from the E.coli systems.
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 4) cell growth assay.
Purified protein is greater than 95% homogeneous based on SDS-PAGE gel analysis. 1 unit equals 1 nanogram of purified protein. 1 unit is sufficient for a gel mobility shift assay in a 20 µl reaction; 50 units are sufficient for reconstituted transcription assay and 100 units are sufficient for a protein-protein interaction assay.
variable in different lots 1x dilution buffer A: 20 mM Tris-Cl (pH 8.0), 20% glycerol, 100 mM KCl, 1 mM DTT and 0.2 mM EDTA
References: 1. Chen, X. et al., (1996) Genes & Dev. 10, 2438-2451 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. Shaw, P. et al., (1996) Oncogene 12, 20393-20802 6. Gu, W., et al., (1997) Cell 90, 595-606 7. Xu et al., (2005) Nat Cell Bio, 7:165-171
DISCLAIMER
This products is recommended For RESEARCH USE ONLY and is Not qualified for Use in Diagnostic or Therapeutic Procedures.
