Description:This HeLa cell nuclear extract is specifically recommended for: 1) in vitro splicing; 2) protein-DNA/RNA and protein-protein interactions; and 3) source of individual splicing factors. Although this extract also contains most transcription factors, it is, however, not recommended for in vitro transcription assays.
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Description | The HeLa cell nuclear extract was prepared as described by Manley et al. (1) and Krainer et al. (2). Although this extract contains all basal transcription factors, as well as most gene-specific activators and cofactors, it is prepared specifically for the purpose of pre-mRNA splicing and polyadenylation (3, 4). Therefore, it has been used as the source of individual polyadenylation factors, splicing factors, and for the cell-free system to study the mechanism of pre-mRNA processing (5-8). | | Source | Extracted from Cervical Tumor cells. | | Concentration | 7.5 mg protein/ml (in 1x dilution buffer B) | | Unit Definition (Activity) | 1-5μl is sufficient for a gel mobility shift assay in a 20μl reaction; 5-10μl is sufficient for in vitro splicing assay and 20-50μl is sufficient for a protein-protein interaction assay. Extract activity has been validated using an in vitro pre-mRNA splicing assay. | | Buffer and Storage | 1x dilution buffer B: 20 mM HEPES-Na (pH 7.9), 20% Glycerol, 42 mM (NH4)2SO4, 0.5 mM DTT and 0.2 mM EDTA.The extract is stored at -80oC. Avoid repeated freezing and thawing cycles. | | Applications | The HeLa cell nuclear extract of this catalog number is specifically recommended for: 1) in vitro splicing; 2) protein-DNA/RNA and protein-protein interactions; and 3) as a source of individual splicing factors. Although this extract also contains most transcription factors, it is, however, not recommended for in vitro transcription assays. Please use our specific Hela Nuclear Extract (Cat# P1001) for the purpose of In Vitro transcription. For Research Use Only. | | References | 1. Manley, J.L., et al., (1980) Proc. Natl. Acad. Sci. USA 77, 5706-5710
2. Krainer, A.R., et al., (1984) Cell 36, 993-1005
3. Kramer, A., et al., (1987) J. Biol. Chem. 262, 17630-17640
4. Takagaki, Y., et al., (1989) Genes Dev. 3, 1711-1724
5. Gerke, V., et al., (1986) Cell 47, 973-984
6. Ruskin, B., et al., (1984) Cell 38, 317-331
7. Rossi, F., et al., (1996) Nature 381, 80-82
8. Krainer, A.R., et al., (1990) Cell 62, 35- |
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