Revision 2
#88503
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For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:
W
Reactivity:
H
Sensitivity:
Endogenous
Source/Isotype:
Rabbit IgG
UniProt ID:
#P62987, #P0CG48, #P0CG47, #P62979
Entrez-Gene Id:
7311, 7316, 7314, 6233
Product Usage Information
Application | Dilution |
---|---|
Western Blotting | 1:1000 |
Storage
Specificity/Sensitivity
Phospho-Ubiquitin (Ser65) (E2J6T) Rabbit mAb (Biotinylated) recognizes endogenous levels of ubiquitin protein only when phosphorylated at Ser65.
Species predicted to react based on 100% sequence homology
Mouse, Rat
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser65 of human ubiquitin protein.
Description
This Cell Signaling Technology® antibody is conjugated to biotin under optimal conditions. The biotinylated antibody is expected to exhibit the same species cross-reactivity as the unconjugated Phospho-Ubiquitin (Ser65) (E2J6T) Rabbit mAb #62802.
Background
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 phosphorylated 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 Parkinson's disease (10,11).
Background References
- Ciechanover, A. (1998) EMBO J 17, 7151-60.
- Hochstrasser, M. (2000) Nat Cell Biol 2, E153-7.
- Hochstrasser, M. (2000) Science 289, 563-4.
- Bernardi, R. et al. (2000) Oncogene 19, 2447-54.
- Aberle, H. et al. (1997) EMBO J 16, 3797-804.
- Salomoni, P. and Pandolfi, P.P. (2002) Nat Cell Biol 4, E152-3.
- Jesenberger, V. and Jentsch, S. (2002) Nat Rev Mol Cell Biol 3, 112-21.
- Kane, L.A. et al. (2014) J Cell Biol 205, 143-53.
- Koyano, F. et al. (2014) Nature 510, 162-6.
- Kitada, T. et al. (1998) Nature 392, 605-8.
- Valente, E.M. et al. (2004) Science 304, 1158-60.
Species Reactivity
Species reactivity is determined by testing in at least one approved application (e.g., western blot).
Western Blot Buffer
IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v nonfat dry milk, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
Applications Key
W: Western Blotting
Cross-Reactivity Key
H: Human M: Mouse R: Rat Hm: Hamster Mk: Monkey Vir: Virus Mi: Mink C: Chicken Dm: D. melanogaster X: Xenopus Z: Zebrafish B: Bovine Dg: Dog Pg: Pig Sc: S. cerevisiae Ce: C. elegans Hr: Horse GP: Guinea Pig Rab: Rabbit G: Goat All: All Species Expected
Trademarks and Patents
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.
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