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ASF/SF2 (SFRS1 or SRP30a from E. Coli )
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
This products is recommended For RESEARCH USE ONLY and is Not qualified for Use in Diagnostic or Therapeutic Procedures.
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