CycLex Proteasome Enrichment Activity Assay Kit
| Code | Size | Price |
|---|
| MBL-CY-7002 | 20 Assays | £524.00 |
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Overview
Regulatory Status: RUO
Shipping:
4°C
Storage:
4°C
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Further Information
Background:
The ubiquitin-proteasome pathway is the principle pathway of proteolysis in eukaryotic cells and may contribute to controlling the intracellular levels of a variety of short-lived proteins (1-3), in addition to degrading abnormal proteins in the cytosol and nucleus. Protein substrates are marked with a poly-ubiquitin chain (4) and then degraded to peptides and free ubiquitin by a large multicatalytic complex, the proteasome, which exists within all eukaryotic cells (1-3). Numerous examples of regulatory proteins have been found to undergo ubiquitin-dependent proteolysis.
Protein substrates of the ubiquitin-proteasome pathway include a number of cell regulatory molecules, such as cyclins, the Myc oncogene protein, and p53, and the regulated degradation of these molecules has been linked to the control of cell proliferation and cell cycle progression (5-7). By controlling the intracellular levels of such proteins, the activity of the ubiquitin-proteasome pathway might also be linked to apoptosis.
A class of proteins containing an N-terminal ubiquitin-like (UbL) domain has been implicated in the regulation of proteolysis (8-13). A member of this superfamily, Saccharomyces cerevisiae Rad23 was first identified for its role in DNA repair (14). For the human homolog of Rad23, two proteins hHR23A (363 residues) and hHR23B (409 residues) exist, which share an identity of 59 % on the amino acid level. Differences in function between the two homologs are unclear (15). hHR23A and hHR23B also contain N-terminal 80 residues, UbL domain that have been shown to interact with the 26S proteasome. Both UbL domain fused to GST was shown to pull down 20S proteasome subunits containing chymotrypsin-like proteolytic activity.
Description:
The CycLex Research Product Proteasome Enrichment & Activity Assay kit is designed to enrich proteasome from cell lysate and measure its protease activity. Since the amino acid sequence of ubiquitin-like domain in hHR23B is well conserved among mammalian, this kit can be used for all mammalian cells.
Kit Components:
UbL-Resin, Control-Resin, 10X Cell Extraction Buffer, Proteasome Substrate (Suc-LLVY-AMC), 10X Assay Buffer
Target:
Proteasome
References
1. Goldberg A. L., Stein R., Adams J. Chem. Biol., 2: 503-508, 1995.
2. Coux O., Tanaka K., Goldberg A. L. Annu. Rev. Biochem., 65: 801-847, 1996.
3. King R. W., Deshaies R. J., Peters J-M., Kirschner M. W. Science, 274: 1652-1659, 1996.
4. Chau V., Tobias J. W., Bachmair A., Marriott D., Ecker D. J., Gonda D. K., Varshavsky A. Science, 243: 1576-1583, 1989.
5. Hochstrasser, M. Curr. Opin. Cell Biol. 7: 215-223, 1995
6. Ciechanover, A. Cell 79: 13-21, 1994
7. Jentsch, S., and Schlenker, S. Cell 82: 881-884, 1995
8. Schauber,C et al. Nature, 391: 715-718, 1998
9. Hiyama,H et al. J. Biol. Chem., 274: 28019-28025, 1999
10. Kleijnen,M.F et al. Mol. Cell, 6: 409-419, 2000
11. Elsasser,S. et al. Nat Cell .Biol., 4: 725-730, 2002
12. Funakoshi,M. et al. Proc. Natl Acad. Sci. USA, 99: 745-750, 2002
13. Rao,H. and Sastry,A. J. Biol. Chem., 277: 11691-11695, 2002
14. Haynes,R.H. and Kunz,B.A The Molecular Biology of the Yeast Saccharomyces. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY, pp. 371-414, 1981
15. Sugasawa,K. et al. Mol. Cell. Biol., 17: 6924-6931, 1997