Cell Signaling Technology
XP Monoclonal Antibody

Product Pathways - Protein Stability

UCHL1 (D8R2I) XP® Rabbit mAb #11896

Applications Reactivity Sensitivity MW (kDa) Isotype
W IF-IC H M R Mk Endogenous 27 Rabbit IgG

Applications Key:  W=Western Blotting  IF-IC=Immunofluorescence (Immunocytochemistry)
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

UCHL1 (D8R2I) XP® Rabbit mAb recognizes endogenous levels of total UCHL1 protein. This antibody does not cross-react with other UCH family members.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Asp110 of human UCHL1 protein.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines and rat brain tissue using UCHL1 (D8R2I) XP® Rabbit mAb (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).

Western Blotting

Western Blotting

Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with constructs expressing Myc/DDK-tagged full-length human UCHL1 (hUCHL1-Myc/DDK; +), Myc/DDK-tagged full-length human UCHL3 (hUCHL3-Myc/DDK; +), Myc/DDK-tagged full-length human UCHL5 (hUCHL5-Myc/DDK; +), and Myc/DDK-tagged full-length human BAP1 (hBAP1-Myc/DDK; +), using UCHL1 (D8R2I) XP® Rabbit mAb (upper) or DYKDDDDK Tag Antibody #2368 (lower).

IF-IC

IF-IC

Confocal immunofluorescent analysis of DU 145 (positive; left) and LNCaP (negative; right) cells using UCHL1 (D8R2I) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).


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 Ubiquitin C-terminal Hydrolase (UCH) family of DUBs, which all possess 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.UCHL1 (PGP 9.5/PARK5) functions as a deubiquitinating enzyme and monoubiquitin stabilizer. In vitro studies have demonstrated that UCHL1 can hydrolyze isopeptide bonds between the C-terminal glycine of Ub and the ε-amino group of lysine on target proteins. UCHL1 is also involved in the cotranslational processing of pro-ubiquitin and ribosomal proteins translated as ubiquitin fusions (5-7). Mice deficient in UCHL1 experience spasticity, suggesting that UCHL1 activity is required for the normal structure and function of the neuromuscular junction (5-7). Research studies have implicated loss of UCHL1 expression in numerous human malignancies, such as prostate, colorectal, renal, and breast carcinomas. Investigators have shown that in breast carcinomas, loss of UCHL1 expression can be attributed to hyper-methylation of the UCHL1 promoter (8). While loss of UCHL1 expression is implicated in human carcinogenesis, mutation of UCHL1 has been implicated in neurodegenerative diseases such as Parkinson's and Alzheimer's (6,7).

  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. Todi, S.V. and Paulson, H.L. (2011) Trends Neurosci 34, 370-82.
  6. Setsuie, R. and Wada, K. (2007) Neurochem Int 51, 105-11.
  7. Day, I.N. and Thompson, R.J. (2010) Prog Neurobiol 90, 327-62.
  8. Xiang, T. et al. (2012) PLoS One 7, e29783.

Application References

Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!

Companion Products

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

Products