| Purification and Quality Control | The His-tag recombinant protein is purified by affinity chromatography in combination with FPLC columns. The purified SC35 is greater than 95% homogeneous based on SDS-PAGE analysis. | | Unit Definition (Activity) | 1 unit equals 1 nanogram of purified protein. 1-5 units are the amount sufficient for a gel mobility shift assay in a 20 µl reaction, 100 units are sufficient for a protein-protein interaction assay and 100-500 units are required for a splicing assay in a 20 μl reaction. | | Applications | Recombinant SC35 protein can be used for: 1) in vitro function studies including pre-mRNA splicing, cross linking and other RNA binding assays; 2) protein-protein interaction assay; and 3) cell growth and proliferation 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 | PR264; SC-35; SC35; SFRS2; SFRS2A; SRp30b, serine/arginine-rich splicing factor 2. | | Protein Sequence | GRPPPDVEGM TSLKVDNLTY RTSPDTLRRV FEKYGRVGDV YIPRDRYTKE SRGFAFVRFH
DKRDAEDAMD AMDGAVLDGR ELRVQMARYG RPPDSHHSRR GPPPRRYGGG GYGRRSRSPR
RRRRSRSRSR SRSRSRSRSR YSRSKSRSRT RSRSRSTSKS RSARRSKSKS SSVSRSRSRS
RSRSRSRSPP PVSKRESKSR SRSKSPPKSP EEEGAVSS | | Background | SC35, a member of the SR protein family, is an essential pre-mRNA splicing factor and can function as an essential splicing factor as well as an alternative splicing factor (1, 2). Phosphorylation of serine residues located within the SR domain directly regulates SC35 activity and compartmentalization of other SR splicing factors (3, 4). In addition to interacting with RNA and other splicing factors, SC35 has been shown to interact either directly or indirectly with the C-terminal domain (CTD) of the largest subunit of RNA polymerase II, thereby suggesting a potential role of SC35 in coordinating transcription and pre-mRNA splicing (5, 6). | | References | 1. Fu, X.D., et al., (1992) Science 256, 535-538
2. Zahler, A.M., et al., (1992) Genes & Dev. 6, 837-847
3. Gui, J.F., et al., (1994) Proc. Natl. Acad. Sci. USA 91, 10824-10828
4. Colwill, K., et al., (1996) EMBO J. 15, 265-275
5. Mortillaro, M.J., et al., (1997) Proc. Natl. Acad. Sci. USA 93, 8253-8257
6. Misteli, T., et al., (1997) Nature 387, 523-527 | | Image of SDS-PAGE /Western-blot |  |
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