| 货号 | 37642S |
| 反应种属 | Human |
| 来源宿主 | Rabbit |
| 应用 | WB,IP |
| 目标/特异性 | Phospho-Ubiquitin (Ser65) Antibody recognizes endogenous levels of ubiquitin protein only when phosphorylated at Ser65. |
| 使用方法 | Western Blotting (1:1000) Immunoprecipitation (1:200) |
| 供应商 | CST |
| 灵敏度 | Endogenous |
| 背景 | Ubiquitin is a conserved polypeptide unit that plays an important role in the ubiquitin-proteasome pathway. Ubiquitin can be covalently linked to many cellular proteins by the ubiquitination process, which targets proteins for degradation by the 26S proteasome. Three components are involved in the target protein-ubiquitin conjugation process. Ubiquitin is first activated by forming a thiolester complex with the activation component E1; the activated ubiquitin is subsequently transferred to the ubiquitin-carrier protein E2, then from E2 to ubiquitin ligase E3 for final delivery to the epsilon-NH2 of the target protein lysine residue (1-3). The ubiquitin-proteasome pathway has been implicated in a wide range of normal biological processes and in disease-related abnormalities. Several proteins such as IκB, p53, cdc25A, and Bcl-2 have been shown to be targets for the ubiquitin-proteasome process as part of regulation of cell cycle progression, differentiation, cell stress response, and apoptosis (4-7). Ubiquitin is phosphorlated at Ser65 by PINK1, leading to activation of the E3 ubiquitin ligase Parkin (8, 9). PINK1 accumulates on depolarized mitochondria, resulting in phosphorylation of ubiquitin and activation of Parkin, which then triggers the mitophagy pathway to clear damaged mitochondria. Loss-of-function mutations in PINK1 and Parkin result in early onset Parkinsons disease (10, 11). |
| 存放说明 | -20C |
| 参考文献 | 1 . Ciechanover, A. (1998) EMBO J 17, 7151-60. 2 . Hochstrasser, M. (2000) Nat Cell Biol 2, E153-7. 3 . Hochstrasser, M. (2000) Science 289, 563-4. 4 . Bernardi, R. et al. (2000) Oncogene 19, 2447-54. 5 . Aberle, H. et al. (1997) EMBO J 16, 3797-804. 6 . Salomoni, P. and Pandolfi, P.P. (2002) Nat Cell Biol 4, E152-3. 7 . Jesenberger, V. and Jentsch, S. (2002) Nat Rev Mol Cell Biol 3, 112-21. 8 . Kane, L.A. et al. (2014) J Cell Biol 205, 143-53. 9 . Koyano, F. et al. (2014) Nature 510, 162-6. 10 . Kitada, T. et al. (1998) Nature 392, 605-8. 11 . Valente, E.M. et al. (2004) Science 304, 1158-60. |
Western blot analysis of extracts from PC-3 cells, untreated (-) or treated with carbonyl cyanide 3-chlorophenylhydrazone (CCCP) (30 μM, indicated times; +), using Phospho-Ubiquitin (Ser65) Antibody (upper), PINK1 (D8G3) Rabbit mAb #6946 (middle), or β-Actin (D6A8) Rabbit mAb #8457 (lower). | |
Immunoprecipitation of phospho-ubiquitin (Ser65) from extracts of PC-3 cells treated with carbonyl cyanide 3-chlorophenylhydrazone (CCCP) (30 μM, 6 hr). Lane 1 is 10% input, lane 2 is precipitated with Normal Rabbit IgG #2729, and lane 3 is Phospho-Ubiquitin (Ser65) Antibody. Western blot was performed using Phospho-Ubiquitin (Ser65) Antibody. |
