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Ubiquitin E3 Ligase Complex Antibody Sampler Kit
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Orders:

877-616-CELL (2355)

[email protected]

Support:

877-678-TECH (8324)

Web:

3 Trask Lane | Danvers | Massachusetts | 01923 | USA

For Research Use Only. Not for Use in Diagnostic Procedures.

Product Includes Product # Quantity Mol. Wt Isotype/Source
CUL4A Antibody269920 µl80, 82 kDaRabbit
CYLD (D6O5O) Rabbit Monoclonal Antibody1279720 µl109 kDaRabbit IgG
DDB-1 (D4C8) Rabbit Monoclonal Antibody699820 µl127 kDaRabbit IgG
DDB-2 (D4C4) Rabbit mAb541620 µl43 kDaRabbit IgG
RBX1 (D3J5I) Rabbit Monoclonal Antibody1192220 µl13 kDaRabbit IgG
Skp1 (D3J4N) Rabbit Monoclonal Antibody1224820 µl19 kDaRabbit IgG
Skp2 (D3G5) Rabbit Monoclonal Antibody265220 µl48 kDaRabbit IgG
beta-TrCP (D12C8) Rabbit Monoclonal Antibody1198420 µl62 kDaRabbit IgG
Anti-rabbit IgG, HRP-linked Antibody7074100 µlGoat

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

Description

The Ubiquitin E3 Ligase Complex Antibody Sampler Kit provides an economical means to study multiple protein components of ubiquitin E3 ligase complexes. The kit includes enough antibody to perform two western blot experiments per primary 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

Ubiquitin can be covalently linked to many cellular proteins by the ubiquitination process, which targets proteins for degradation by the 26S proteasome. Ubiquitin is first activated by forming a thiolester complex with the activation component E1. The activated ubiquitin is subsequently transferred to the ubiquitin-carrier protein E2, and then from E2 to ubiquitin ligase E3 for final delivery to the ε-NH2 of the target protein lysine residue (1-3). Research studies suggest that activated E2 associates transiently with E3, and the dissociation is a critical step for ubiquitination (4).

S phase kinase-associated protein 1 (Skp1) is a critical scaffold protein of the Skp1/CUL1/F-box (SCF) E3 ubiquitin ligase protein complex. Various F-box proteins (e.g. β-TrCP, Skp2) mediate an interaction with Skp1 via their defining and conserved domain of 40 amino acids and with substrates to be ubiquitinated (5). RING-box protein 1 (RBX1 or ROC1) is another essential component of the SCF complex (6). RBX1 mediates the neddylation of CUL1, which activates SCF E3 ligase by facilitating the ubiquitin transfer from E2 to substrates (7-9). The RING finger domain of RBX1 is required for ubiquitin ligation (10).

Cullin-4 (CUL4) is a member of the cullin family of related ubiquitin ligases (11). The carboxy-terminal domain of CUL4 interacts with Rbx1 and E2 enzyme while the amino-terminal CUL4 domain interacts with BPB domain of UV-damaged DNA binding protein DDB-1 to form a CUL4-DDB1 ubiquitin ligase complex (12). Damaged DNA-Binding Protein (DDB) consists of a 127 kDa subunit (DDB-1) and a 48 kDa subunit (DDB-2) that contribute to the formation of the UV-damaged DNA-binding protein complex (UV-DDB) (13-15). In conjunction with CUL4A and RBX1, the UV-DDB complex forms an E3 ubiquitin ligase that recognizes a broad spectrum of DNA lesions. The complex polyubiquitinates components of the nucleotide excision repair pathway (16-18).

Background References

  1. Ciechanover, A. (1998) EMBO J 17, 7151-60.
  2. Hochstrasser, M. (2000) Nat Cell Biol 2, E153-7.
  3. Hochstrasser, M. (2000) Science 289, 563-4.
  4. Deffenbaugh, A.E. et al. (2003) Cell 114, 611-22.
  5. DeSalle, L.M. and Pagano, M. (2001) FEBS Lett 490, 179-89.
  6. Zheng, N. et al. (2002) Nature 416, 703-9.
  7. Kamura, T. et al. (1999) Genes Dev 13, 2928-33.
  8. Morimoto, M. et al. (2003) Biochem Biophys Res Commun 301, 392-8.
  9. Pan, Z.Q. et al. (2004) Oncogene 23, 1985-97.
  10. Sun, Y. et al. (2001) Antioxid Redox Signal 3, 635-50.
  11. Petroski, M.D. and Deshaies, R.J. (2005) Nat Rev Mol Cell Biol 6, 9-20.
  12. Lee, J. and Zhou, P. (2007) Mol Cell 26, 775-80.
  13. Reardon, J.T. et al. (1993) J Biol Chem 268, 21301-8.
  14. Keeney, S. et al. (1993) J Biol Chem 268, 21293-300.
  15. Hwang, B.J. and Chu, G. (1993) Biochemistry 32, 1657-66.
  16. Chu, G. and Chang, E. (1990) Proc Natl Acad Sci U S A 87, 3324-7.
  17. Hirschfeld, S. et al. (1990) Mol Cell Biol 10, 2041-8.
  18. Payne, A. and Chu, G. (1994) Mutat Res 310, 89-102.

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Limited Uses

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Orders: 877-616-CELL (2355) • [email protected] • Support: 877-678-TECH (8324) • [email protected] • Web: cellsignal.com
For Research Use Only. Not for Use in Diagnostic Procedures.