| Purification and Quality Control | The GST-fusion RAP30 is purified by affinity chromatography and/or other chromatographic means on FPLC system. The purified GST-RAP30 is greater than 90% homogeneous based on SDS-PAGE analysis. | | Unit Definition (Activity) | 1 unit equals 1 nanogram of purified protein. 100 units (ng) are sufficient for protein-protein interaction assays. | | Applications | RAP30 can interact with RNA polymerase II, with TFIIB, and with DNA (3-5). GST-RAP30 has been used in protein-protein interactions assay (6). For Research Use Only. | | Formulation and Storage | The protein is in 20 mM Tris-Cl (pH 8.0), 20% Glycerol, 100 mM KCl, 1 mM DTT and 0.2 mM EDTA. Stored at -70˚C before use. Avoid repeated freeze thaw cycles. | | Synonym | | | Protein Sequence | | | Background | The transcription factor IIF (TFIIF) is composed of 58 kDa (RAP74) and 26 kDa (RAP30) subunits as a heterodimer, and was first identified through the ability to interact with immobilized RNA polymerase II (1, 2). The RAP30 subunit of TFIIF contains two distinct regions with sequence similarity to E coli factors and can deliver RNA polymerase II to the promoter to support transcription initiation in the absence of RAP74 (2, 3). | | References | 1. Sopta, M. et al., (1985) J. Biol. Chem. 260, 10353-10360
2. Sopta, M. et al., (1989) Nature 341, 410-414
3. McCracken, S. et al., (1991) Science 253, 900-902
4. Ha, I. Et al., (1993) Genes & Dev. 7, 1021-1032
5. Tan, S. et al., (1994) Proc. Natl. Acad. Sci. USA 92, 6042-6046
6. Kaludov, N., et al., (2000) Nucleic Acids Res. 28, 1921-1928 | | Image of SDS-PAGE /Western-blot | |
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