| Description | The TATA-binding protein (TBP) is believed to function as an essential factor of the general transcription machinery and to be involved in transcription by all three eukaryotic RNA polymerases (pol I, II, and III).
TBP specifically binds to the TATA element at the promoter region and interacts with numerous transcription factors, including TBP-associated factors (TAFs), activators, and some tumor suppressor proteins (1-3).
The His tagged recombinant TBP is isolated from an E. coli strain that carries the coding sequence for human TBP under the control off T7 promoter (3). | | Target gene sequence | MDQNNSLPPYAQGLASPQGAMTPGIPIFSPMMPYGTGLTPQPIQ NTNSLSILEEQQRQQQQQQQQQQQQQQQQQQQQQQQQQQQQQQQQQQQQQQAVAAAAV QQSTSQQATQGTSGQAPQLFHSQTLTTAPLPGTTPLYPSPMTPMTPITPATPASESSG IVPQLQNIVSTVNLGCKLDLKTIALRARNAEYNPKRFAAVIMRIREPRTTALIFSSGK MVCTGAKSEEQSRLAARKYARVVQKLGFPAKFLDFKIQNMVGSCDVKFPIRLEGLVLT HQQFSSYEPELFPGLIYRMIKPRIVLLIFVSGKVVLTGAKVRAEIYEAFENIYPILKG FRKTT | | Formulation and storage | Protein is stored in 20 mM Tris-Cl (pH 8.0), 20% Glycerol, 100 mM KCl, 1 mM DTT and 0.2 mM EDTA buffer and kept at -80°C. Avoid repeatly freeze thaw cycles. | | Applications | TBP can be used for 1) in vitro transcription; 2) gel mobility shift assay in the presence of double-stranded oligonucleotide containing a TATA element; 3) for in vitro footprinting assay in the presence of TATA-containing DNA fragment (1, 4) and 4) for protein-protein interactions assay. 1 unit equals 1 nanogram of purified protein. 1 unit is the amount sufficient for a gel mobility shift assay in a 20 µl reaction. 10 units are sufficient for a 25 µl reconstituted transcription reaction and 100 units are sufficient for a protein-protein interaction assay detected by immuno-blot system. | | Synonyms | GTF2D; GTF2D1; HDL4; MGC117320; MGC126054; MGC126055; SCA17; TFIID | | References | 1. Horikoshi, M. et al., (1988) Cell 54, 1033-1042
2. Hernandez, N. (1993) Genes & Dev. 7, 1291-1308
3. Holffman, A. et al., (1991) Nucleic Acids Res. 19, 6337-6338
4. Ge, H. et al., (1994) J. Biol. Chem. 269, 17136-17140
5. Grace et al., (2004) J. Bio Chem, 279:8102-8110
6. Francois et al., (2005) J. Virology, 79:17, 11082-11094, | | SDS-PAGE with Coomassie Blue staining |  |
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