Cell Signaling Technology

Product Pathways - Protein Stability

UCHL1 (D8R2I) XP® Rabbit mAb #11896

Deubiquitinating Enzymes   PGP 9.5   PGP9.5   UCHL-1   UCHL1  

No. Size Price
11896S 100 µl ( 10 western blots ) ¥3,750.00 现货查询 购买询价
11896 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human,Mouse,Rat,Monkey, Endogenous 27 Rabbit IgG
IF-IC 1:50

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting, IF-IC=Immunofluorescence (Immunocytochemistry),

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.

UCHL1 (D8R2I) XP® Rabbit mAb兔单抗内源性的UCHL1总蛋白。该抗体与UCH家族的其他成员无交叉反应性。

Source / Purification

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

该单克隆抗体经合成与人UCHL1蛋白的Asp110邻近氨基酸残基序列一致的肽段,免疫动物产生。

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).Confocal immunofluorescent检测DU 145 细胞(阳性; 左) 和LNCaP细胞(阴性;右),采用UCHL1 (D8R2I) XP® Rabbit mAb(绿色)。内参照采用DY-554 phalloidin标记(红色)。

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 blot检测多种细胞提取物和大鼠大脑组织,采用UCHL1 (D8R2I) XP® Rabbit mAb (upper)或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).Western blot检测mock transfected (-)或转染表达Myc/DDK-tagged人全长UCHL1 (hUCHL1-Myc/DDK; +)、Myc/DDK-tagged人全长UCHL3 (hUCHL3-Myc/DDK; +)、Myc/DDK-tagged人全长UCHL5 (hUCHL5-Myc/DDK; +)、Myc/DDK-tagged人全长BAP1 (hBAP1-Myc/DDK; +)的293T细胞,采用UCHL1 (D8R2I) XP® Rabbit mAb (upper)或者DYKDDDDK Tag Antibody #2368 (lower)。

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 carboxy-terminal hydrolase (UCH) family of DUBs, which all possess a conserved catalytic UCH domain of about 230 amino acids. UCHL5 and BAP1 have unique, extended carboxy-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 carboxy-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 neuromuscular junction structure and function (5-7). Research studies have described loss of UCHL1 expression in numerous human malignancies, such as prostate, colorectal, renal, and breast carcinomas. Investigators have shown that loss of UCHL1 expression in breast carcinomas can be attributed to hyper-methylation of the UCHL1 gene 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).

蛋白泛素化和去泛素化作用是由泛素酶(UBEs)和去泛素酶(DUBs)催化的可逆过程(1,2)。DUBs分为5个亚家族:USP, UCH, OTU, MJD和JAMM 。UCHL1, UCHL3, UCHL5/UCH37和BRCA-1-相关蛋白-1(BAP1)属于DUBs的泛素碳末端的水解酶(UCH)家族,它们均具有含有约230个氨基酸的保守的催化结构域(UCH结构域)。UCHL5和BAP1有独特的延长的碳末端尾。UCHL1在神经组织和睾丸中大量表达,而UCHL3的表达则广泛分布(3,4)。尽管UCHL1和UCHL3属于亲近的UCH家族,并具有大约53%的同源性,但它们的生化特性却不同,UCHL1结合单泛素,UCHL3则显示则对泛素和泛素样分子NEDD8显示双重特性。UCHL1 (PGP 9.5/PARK5)作为去泛素化酶和单泛素稳定剂。体外研究表明,UCHL1能够水解Ub羧基末端甘氨酸和靶蛋白上的赖氨酸ε-氨基团之间的异肽键。UCHL1也在泛素前和核糖体蛋白翻译为泛素融合物的翻译过程中起作用(5-7)。UCHL1缺陷的小鼠呈现痉挛状态,表明UCHL1的活性对于正常结构和神经肌肉连接的功能是必须的(5-7)。研究表明在许多人恶性肿瘤中缺乏UCHL1的表达,诸如前列腺、结直肠、肾和乳腺癌。研究者已经证实乳腺癌,缺乏UCHL1的表达,有助于UCHL1启动子的超甲基化(8)。尽管人恶性肿瘤中缺乏UCHL1的表达,在神经变性疾病如Parkinson's 和Alzheimer's中均可能存在UCHL1基因的突变(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. 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.

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