| Purification and Quality Control | The His-tag biotinylated recombinant protein is purified by affinity chromatography in combination with FPLC columns. Then His-tag was removed by protease. Purified biotinylated SRC1-RID is greater than 90% homogeneous based on SDS-PAGE analysis. | | Unit Definition (Activity) | 1 unit equals 1 nanogram of purified protein. 100 units are sufficient for a protein-protein interaction assay. | | Applications | The biotinylated SCR1-RID (627-786) can be used for protein-protein interaction assays, particularly suitable for high throughput screening (HTS) assays. | | 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 | bHLHe42; bHLHe74; F-SRC-1; KAT13A; MGC129719; MGC129720; RIP160; SRC1 and Homo sapiens nuclear receptor coactivator 1 (NCOA1). | | Protein Sequence | SQTSHKLVQL LTTTAEQQLR HADIDTSCKD VLSCTGTSNS ASANSSGGSC PSSHSSLTER
HKILHRLLQE GSPSDITTLS VEPDKKDSAS TSVSVTGQVQ GNSSIKLELD ASKKKESKDH
QLLRYLLDKD EKDLRSTPNL SLDDVKVKVE KKEQMDPCNT | | Background | Steroid receptor coactivator 1 (SRC1) is a transcriptional coactivoator that mediates the activating functions of many of the nuclear hormone receptors. It is also known as NCoA1 and is a member of the SRC/p160 coactivator family (1). SRC1 has been shown to be over expressed in some cancers (2). SRC1 is a 160 kDa protein that contains several LXXLL motifs, which are involved in nuclear receptor interaction (3). The region 627-786 contains 3 LXXLL motifs that are involved in interaction with nuclear hormone receptors (4,5) and has been previously used in assays detecting ligand-dependent receptor-cofactor interactions (6). This protein does not contain His tag and will be useful for assays where a native untagged protein is desired. Lane 1 corresponds to 6His-tagged and biotinylated SRC-RID; Lane 2 corresponds to biotinylated SRC-RID without 6His tag. | | References | 1. Onate, S. et al., (1995) Science 270, 1354-1357
2. Yanase, T., et al., (2004) Intern. Med. 43, 368-373
3. Heery, D. et al., (1997) Nature 387, 733-736
4. Xu, J. and Li, Q., (2003) Mol. Endocrinol. 17, 1681-1692
5. Bai, Y. and Giguere, V., (2003) Mol. Endocrinol. 17, 589-599
6. Cho, M. et al., (2003) J. Biochem. Mol. Biol. 36, 207-213 |
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