MY ACCOUNT
GP53-PACK
P 53 Package contains
- full length p53 (Cat# p2001-01, 5ug)
- C-terminal deleted p53(1-363; Cat# p2002-01, 5ug)
- p53(1-343; Cat# p2003-01, 5ug)
- 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.
