| Purification and Quality Control | The His-tag recombinant protein is purified by affinity chromatography in combination with FPLC columns. The purified P300-Ch1 is greater than 90% homogeneous based on SDS-PAGE analysis. |
| Unit Definition (Activity) | 1 unit equals 1 nanogram of purified protein. 1-5 units are sufficient for a gel mobility shift assay in a 20µl reaction; 50-100 units are sufficient for reconstituted transcription assay and 100-200 units are sufficient for a protein-protein interaction assay or an acetylation assay. |
| Applications | Recombinant p300-Ch1 can be used for: 1) protein-protein interaction assay; 2) in vitro transcription assay; 3) in vitro acetylation assay; and 4) cell growth assay. For Research Use Only. |
| 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. |
| Protein Sequence | MGQQPAPQVQ QPGLVTPVAQ GMGSGAHTAD PEKRKLIQQQ LVLLLHAHKC QRREQANGEV
RQCNLPHCRT MKNVLNHMTH CQSGKSCQVA HCASSRQIIS HWKNCTRHDC PVCLPLKNAG
DKRNQQPILT GAPVGLGNPS SLGVGQQSAP NLSTVSQIDP SSIERAYAAL GLPYQVNQMP
TQPQVQAKNQ QNQQPGQSPQ GMRPMSNMSA SPMGVNGGVG VQTPSLLSDS |
| 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. demonstrated a novel mechanism of targeted p300 regulation of gene expression (5-7). The Zn(2+)-binding cysteine/histidine-rich 1 (CH1) domain of p300/CBP binds many of these transcription factors. |
| References | 1. Stein, RW. et al., (1990) J. Virol. 64, 4421-4427
2. Chrivia, JC. et al., (1993) Nature 365, 855-859
3. Eckner, R. et al., (1994) Genes & Dev. 8, 869-884
4. Kung, AL. et al., (2000) Genes & Dev. 14, 272-277
5. Imhof, A. et al., (1997) Curr. Biol. 7, 689-692
6. Gu, W. et al., (1997) Cell 90, 595-606
7. Martinez-Balbas, MA. et al., (2000) EMBO J. 19, 662-671 |
| Image of SDS-PAGE /Western-blot |  |
