| Purification and Quality Control | The His-tag recombinant protein is purified by affinity chromatography in combination with FPLC columns. The purified ASF/SF2 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 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 ASF/SF2 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 | ASF; FLJ53078; MGC5228; SF2; SF2p33; SFRS1 and SRp30a. | | Protein Sequence | SGGGVIRGPA GNNDCRIYVG NLPPDIRTKD IEDVFYKYGA IRDIDLKNRR GGPPFAFVEF
EDPRDAEDAV YGRDGYDYDG YRLRVEFPRS GRGTGRGGGG GGGGGAPRGR YGPPSRRSEN
RVVVSGLPPS GSWQDLKDHM REAGDVCYAD VYRDGTGVVE FVRKEDMTYA VRKLDNTKFR
SHEGETAYIR VKVDGPRSPS YGRSRSRSRS RSRSRSRSNS RSRSYSPRRS RGSPRYSPRH
SRSRSRT | | Background | ASF or called SF2, a member of the SR protein family, is an essential pre-mRNA splicing factor required for both single and alternative splicing (1-3). Phosphorylation on serine residues located within the SR domain directly regulates ASF/SF2 activity and compartmentalization of other SR splicing factors (4, 5). In addition to interacting with RNA and other splicing factors, such as U1-70K, U2AF and other SR proteins, ASF/SF2 also directly or indirectly interacts with HIV regulatory protein Rev, the C-terminal domain (CTD) of the largest subunit of RNA polymerase II, and numerous transcription factors, thereby suggesting a potential role of ASF/SF2 in coordinating of transcription and pre-mRNA splicing (6, 7). | | References | 1. Ge, H., et al., (1990) Cell 62, 25-34; (1991) Cell 66, 373-382
2. Krainer, A.R., et al., (1990) Cell 62, 35-42; (1991) Cell 66, 383-394
3. Zahler, A.M., et al., (1992) Genes & Dev. 6, 837-847
4. Gui, J.F., et al., (1994) Proc. Natl. Acad. Sci. USA 91, 10824-10828
5. Colwill, K., et al., (1996) EMBO J. 15, 265-275
6. Tange, T.O., et al., (1996) J. Biol. Chem. 271, 10066-10072
7. Mortillaro, M.J., et al., (1997) Proc. Natl. Acad. Sci. USA 93, 8253-8257
8. Benz et al., (2005) Blood, Vol.105, No.5,2146-2153 |
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