| 货号 | 12415S |
| 描述 | from Cell Signaling Technology (CST) allows the researcher to specifically inhibit UCHL1 expression using RNA interference, a method whereby gene expression can be selectively silenced through the delivery of double stranded RNA molecules into the cell. All SignalSilence® siRNA products from CST are rigorously tested in-house and have been shown to reduce target protein expression by western analysis. |
| 反应种属 | Human |
| 应用 | TFN |
| 供应商 | CST |
| 背景 | 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 Parkinsons and Alzheimers (6,7). |
| 存放说明 | -20C |
| 参考文献 | Nijman, S.M. et al. (2005) Cell 123, 773-86. Nalepa, G. et al. (2006) Nat Rev Drug Discov 5, 596-613. Leroy, E. et al. (1998) Nature 395, 451-2. Kurihara, L.J. et al. (2001) Hum Mol Genet 10, 1963-70. Todi, S.V. and Paulson, H.L. (2011) Trends Neurosci 34, 370-82. Setsuie, R. and Wada, K. Neurochem Int 51, 105-11. Day, I.N. and Thompson, R.J. (2010) Prog Neurobiol 90, 327-62. Xiang, T. et al. (2012) PLoS One 7, e29783. |
Western blot analysis of extracts from 293T cells, transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-), SignalSilence® UCHL1 siRNA I # 12353 (+), or SignalSilence® UCHL1 siRNA II (+), using UCHL1 (D8R2I) XP® Rabbit mAb #11896 (upper) or GAPDH (D16H11) XP® Rabbit mAb #5174 (lower). The UCHL1 (D8R2I) XP® Rabbit mAb confirms silencing of UCHL1 expression, while the GAPDH (D16H11) XP® Rabbit mAb is used as a loading control.Western blot检测转染有100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-)，SignalSilence® UCHL1 siRNA I # 12353 (+)或SignalSilence® UCHL1 siRNA II (+)的293T细胞, 采用UCHL1 (D8R2I) XP® Rabbit mAb #11896 (upper)或GAPDH (D16H11) XP® Rabbit mAb #5174 (lower)。UCHL1 (D8R2I) XP® Rabbit mAb证实了UCHL1的沉默表达，采用GAPDH (D16H11) XP® Rabbit mAb作为内参照。 |
