Revision 1

#4889Store at -20C

1 Kit

(8 x 20 microliters)

Cell Signaling Technology

Orders: 877-616-CELL (2355) [email protected]

Support: 877-678-TECH (8324)

Web: [email protected] cellsignal.com

3 Trask LaneDanversMassachusetts01923USA
For Research Use Only. Not for Use in Diagnostic Procedures.
Product Includes Product # Quantity Mol. Wt Isotype/Source
Skp1 Antibody 2156 20 µl 19 kDa Rabbit 
Skp2 (D3G5) XP® Rabbit mAb 2652 20 µl 48 kDa Rabbit IgG
ISG15 (22D2) Rabbit mAb 2758 20 µl 15 kDa Rabbit IgG
NEDD8 (19E3) Rabbit mAb 2754 20 µl 9 kDa Rabbit IgG
Ubiquitin (P4D1) Mouse mAb 3936 20 µl Mouse IgG1
UBC3 Antibody 4997 20 µl 32 kDa Rabbit 
SUMO-1 Antibody 4930 20 µl Rabbit 
SUMO-2/3 (18H8) Rabbit mAb 4971 20 µl Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl Goat 
Anti-mouse IgG, HRP-linked Antibody 7076 100 µl Horse 

Please visit cellsignal.com for individual component applications, species cross-reactivity, dilutions, protocols, and additional product information.

Description

This sampler kit provides an economical means to investigate protein folding and stability. The kit contains primary and secondary antibodies to perform two Western blots with each antibody.

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

Background

The small regulatory protein ubiquitin is often covalently linked to many cellular proteins, labeling these targeted proteins for proteasome-mediated degradation. Ubiquitin is first activated by forming a thiolester complex with the E1 activation component. Activated ubiquitin is subsequently transferred to the ubiquitin-carrier protein E2, and then to an E3 ubiquitin ligase for final delivery to the ε-NH2 of the target protein lysine residue (1). The ubiquitin-proteasome pathway has been implicated in a wide range of both normal biological processes and diseases (2,3).
The ubiquitin-like protein family contains three small ubiquitin-related modifier proteins (SUMO-1, -2 and -3), neural precursor cell-expressed developmentally down-regulated protein 8 (NEDD8) and interferon-stimulated 15 kDa protein (ISG15) (4-6). Their covalent attachment to target proteins is a reversible, multi-step process that is analogous to protein ubiquitination. Mature molecules are linked to the activating enzyme E1, conjugated to E2 and ligated to the target proteins by E3 (7-10). Ubiquitin is the predominant regulator for the degradation of a wide range of target proteins (8) while SUMO, NEDD8 and ISG15 modify a limited set of substrates to regulate various other biological processes (4, 11-18).
During ubiquitination, the combinatorial interaction of different E2 and E3 proteins produces variable substrate specificity (4). UBC3 and UBC3B are E2 ubiquitin-carrier proteins (19, 20). The SCF (Skp1/CUL1/F-box) E3 ubiquitin ligase protein complex is composed of three protein components, including the S phase kinase associated protein 1 (Skp1), Cullin homolog 1 (CUL1) and the Skp2 F-box protein (21-23).

  1. Ciechanover, A. (1998) EMBO J. 17, 7151-7160.
  2. Bernardi, R. et al. (2000) Oncogene 19, 2447-2454.
  3. Jesenberger, V. and Jentsch, S. (2002) Nat. Rev. Mol. Cell Biol. 3, 112-121.
  4. Schwartz, D.C. and Hochstrasser, M. (2003) Trends Biochem. Sci. 28, 321-328.
  5. Chiba, T. and Tanaka, K. (2004) Curr. Protein Pept. Sci. 5, 177-184.
  6. Ritchie, K.J. and Zhang, D.E. (2004) Semin. Cell Dev. Biol. 15, 237-246.
  7. Kim, K.I. et al. (2002) J. Cell Physiol. 191, 257-268.
  8. Osaka, F. et al. (1998) Genes Dev. 12, 2263-2268.
  9. Loeb, K.R. and Haas, A.L. (1992) J. Biol. Chem. 267, 7806-7813.
  10. Zhao, C. et al. (2005) Proc. Natl. Acad. Sci. USA 102, 10200-10205.
  11. Matunis, M.J. et al. (1996) J. Cell Biol. 135, 1457-1470.
  12. Duprez, E. et al. (1999) J. Cell Sci. 112 ( Pt 3), 381-393.
  13. Gostissa, M. et al. (1999) EMBO J. 18, 6462-6471.
  14. Rodriguez, M.S. et al. (1999) EMBO J. 18, 6455-6461.
  15. Desterro, J.M. et al. (1998) Mol. Cell 2, 233-239.
  16. Stickle, N.H. et al. (2004) Mol. Cell. Biol. 24, 3251-3261.
  17. Xirodimas, D.P. et al. (2004) Cell 118, 83-97.
  18. Hamerman, J.A. et al. (2002) J. Immunol. 168, 2415-2423.
  19. Semplici, F. et al. (2002) Oncogene 21, 3978-3987.
  20. Pagano, M. et al. (1995) Science 269, 682-685.
  21. Yu, Z.K. et al. (1998) Proc. Natl. Acad. Sci. USA 95, 11324-11329.
  22. Pagano, M. (2004) Mol. Cell 14, 414-416.
  23. Reed, S.I. (2003) Nat. Rev. Mol. Cell Biol. 4, 855-864.

Background References

    Trademarks and Patents

    Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
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