Cell Signaling Technology

Product Pathways - Protein Stability

USP14 (D8Q6S) Rabbit mAb #11931

Applications Reactivity Sensitivity MW (kDa) Isotype
W IP H M R (Hm) (B) (Dg) Endogenous 60 Rabbit IgG

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

Protocols

Specificity / Sensitivity

USP14 (D8Q6S) Rabbit mAb recognizes endogenous levels of total USP14 protein. Based upon sequence alignment, this antibody is predicted to react with both isoform a and isoform b of USP14.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the carboxy terminus of human USP14 protein.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using USP14 (D8Q6S) Rabbit mAb.

Western Blotting

Western Blotting

Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing DDK-tagged full-length human USP14 isoform a (hUSP14-DDK; +), using USP14 (D8Q6S) Rabbit mAb.

IP

IP

Immunoprecipitation of USP14 from 293T cell extracts using Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (lane 2) or USP14 (D8Q6S) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using USP14 (D8Q6S) Rabbit mAb.


Background

Ubiquitinating enzymes (UBEs) catalyze protein ubiquitination, a reversible process countered by deubiquitinating enzyme (DUB) action (1,2). Five DUB subfamilies are recognized, including the USP, UCH, OTU, MJD, and JAMM enzymes. In humans, there are three proteasomal DUBs: PSMD14 (POH1/RPN11), UCH37 (UCH-L5), and Ubiquitin-Specific Protease 14, which is also known as the 60 kDa subunit of tRNA-guanine transglycosylase (USP14/TGT60 kDa). USP14 is recruited to the proteasome through its reversible association with the PSMD2 (S2/hRPN1) subunit of the 19S regulatory particle. Whereas PSMD14 appears to promote substrate degradation (3,4), USP14 is thought to antagonize substrate degradation (5-8). While the underlying mechanism for the opposing roles of these two proteasomal DUBs is still uncertain, it is thought that USP14 removes ubiquitin from substrate upon docking of the substrate with the 26S proteasome. Furthermore, USP14 trims ubiquitin residues from the distal end of the polyubiquitin chain, thus decreasing the affinity of the chain for the ubiquitin receptors of the proteasome, and allowing for enhanced substrate stability (6,9,10). Studies have elucidated a physiologic role for USP14 in regulating synaptic activity in mammals (11). Research studies have shown that targeting this activity with small molecule inhibitors has potential benefits for the treatment of neurodegenerative diseases and cancer (5,12).

  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. Verma, R. et al. (2002) Science 298, 611-5.
  4. Yao, T. and Cohen, R.E. (2002) Nature 419, 403-7.
  5. Lee, B.H. et al. (2010) Nature 467, 179-84.
  6. Lam, Y.A. et al. (1997) Nature 385, 737-40.
  7. Koulich, E. et al. (2008) Mol Biol Cell 19, 1072-82.
  8. Jacobson, A.D. et al. (2009) J Biol Chem 284, 35485-94.
  9. Hanna, J. et al. (2006) Cell 127, 99-111.
  10. Thrower, J.S. et al. (2000) EMBO J 19, 94-102.
  11. Wilson, S.M. et al. (2002) Nat Genet 32, 420-5.
  12. D'Arcy, P. et al. (2011) Nat Med, Epub ahead of print.

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