| Purification and Quality Control | The Flag-tag recombinant protein is purified by affinity chromatography in combination with FPLC columns. The purified p300 is greater than 90% homogeneous based on SDS-PAGE analysis. | | Unit Definition (Activity) | 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. | | Applications | Recombinant p300 can be used for: 1) protein-protein interaction assay; 2) in vitro transcription assay; 3) in vitro acetylation assay; and 4) cell growth assay. | | Formulation and Storage | The protein is in 20mM Tris-HCl pH7.9,100mM NaCl, 0.2mM EDTA, 1mM DTT and 20% glycerol. Stored at -70°C before use. Avoid repeated freeze thaw cycles. | | Synonym | Homo sapiens E1A binding protein p300 (EP300), KAT3B; p300 and RSTS2. | | Background | Human p300 and CBP (CREB binding protein) are highly related transcriptional coactivators. Both proteins have been identified through protein interaction assays (1-3). In addition to interacting with a variety of cellular factors and onco-proteins, loss of the wild type CBP alleles in isolated tumors suggests that CBP/p300 might serve as tumor suppressors (4). The ability of p300 to acetylate many transcription factors, including p53, E2F, TFIIE, and TFIIF etc. demonstrates a novel mechanism of targeted p300 regulation of gene expression (5-7). | | References | 1. Pedeux et. al., Mol Cell Bio, 25:15. 6639-6648, August 2005.
2. Martinez-Balbas, MA. et al., (2000) EMBO J. 19, 662-671
3. Kung, AL. et al., (2000) Genes & Dev. 14, 272-277
4. Imhof, A. et al., (1997) Curr. Biol. 7, 689-692
5. Gu, W. et al., (1997) Cell 90, 595-606
6. Eckner, R. et al., (1994) Genes & Dev. 8, 869-884
7. Chrivia, JC. et al., (1993) Nature 365, 855-859
8. Stein, RW. et al., (1990) J. Virol. 64, 4421-4427 | | Image of SDS-PAGE /Western-blot |  |
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