Cell Signaling Technology

Product Pathways - Protein Stability

UCHL3 (D25E6) Rabbit mAb #8141

Applications Reactivity Sensitivity MW (kDa) Isotype
W H M R (B) (Dg) (Hr) Endogenous 27 Rabbit IgG

Applications Key:  W=Western Blotting
Reactivity Key:  H=Human  M=Mouse  R=Rat  B=Bovine  Dg=Dog  Hr=Horse
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

UCHL3 (D25E6) Rabbit mAb recognizes endogenous levels of total UCHL3 protein. This antibody does not cross-react with either UCHL1 or BAP1 and is not predicted to cross-react with UCHL5.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the amino terminus of human UCHL3 protein.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using UCHL3 (D25E6) Rabbit mAb.

Western Blotting

Western Blotting

Western blot analysis of extracts from 293T cells, mock-transfected (-) or transfected with a Myc/DDK-tagged cDNA expression construct encoding full-length human UCHL3 (hUCHL3, +), using UCHL3 (D25E6) Rabbit mAb.

Background

Protein ubiquitination and deubiquitination are reversible processes catalyzed by ubiquitinating enzymes (UBEs) and deubiquitinating enzymes (DUBs) (1,2). DUBs are categorized into 5 subfamilies: USP, UCH, OTU, MJD, and JAMM. UCHL1, UCHL3, UCHL5/UCH37, and BRCA-1-associated protein-1 (BAP1) belong to the UCH family of DUBs, which all posses a conserved catalytic domain (UCH domain) of about 230 amino acids. UCHL5 and BAP1 have unique extended C-terminal tails. UCHL1 is abundantly expressed in neuronal tissues and testes, while UCHL3 expression is more widely distributed (3,4). Although UCHL1 and UCHL3 are the most closely related UCH family members with about 53% identity, their biochemical properties differ in that UCHL1 binds monoubiquitin and UCHL3 shows dual specificity toward both ubiquitin (Ub) and NEDD8, a Ub-like molecule. In particular, UCHL3 functions as a Ub hydrolase involved in the processing of both Ub precursors and ubiquitinated substrates, generating free monomeric Ub. This is accomplished through the ability of UCHL3 to recognize and hydrolyze isopeptide bonds at the C-terminal glycine of either Ub or NEDD8 (5-7). Recent functional studies have identified UCH-L3 as a critical regulator of adipogenesis through its ability to promote IGF-IR and insulin receptor signaling (8). Furthermore, UCHL3 has been shown to promote deubiquitination, recycling, and cell surface expression of the epithelial sodium channel (9).

  1. Nijman, S.M. et al. (2005) Cell 123, 773-86.
  2. Nalepa, G. et al. (2006) Nat Rev Drug Discov 5, 596-613.
  3. Leroy, E. et al. (1998) Nature 395, 451-2.
  4. Kurihara, L.J. et al. (2001) Hum Mol Genet 10, 1963-70.
  5. Osaka, H. et al. (2003) Hum Mol Genet 12, 1945-58.
  6. Wada, H. et al. (1998) Biochem Biophys Res Commun 251, 688-92.
  7. Kwon, J. (2007) Exp Anim 56, 71-7.
  8. Suzuki, M. et al. (2009) Endocrinology 150, 5230-9.
  9. Butterworth, M.B. et al. (2007) J Biol Chem 282, 37885-93.

Application References

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For Research Use Only. Not For Use In Diagnostic Procedures.

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