Product Pathways - Screening Technologies
K63-linkage Specific Polyubiquitin (D7A11) Rabbit mAb #5621
|5621S||100 µl (10 western blots)||---||In Stock||---|
|5621||carrier free and custom formulation / quantity||email request|
|W||1:1000||All Species Expected||Endogenous||Rabbit IgG|
Species cross-reactivity is determined by western blot.
Applications Key: W=Western Blotting
Specificity / Sensitivity
K63-linkage Specific Polyubiquitin (D7A11) Rabbit mAb detects polyubiquitin chains formed by Lys63 residue linkage. It does not react with monoubiquitin or polyubiquitin chains formed by linkage to a different lysine residue.
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding the Lys63 branch of the human diubiquitin chain.
Western blot analysis of extracts from HeLa cells, untreated or treated with the proteasome inhibitor MG132 (10 µM for 6 hours), using K63-linkage Specific Polyubiquitin (D7A11) Rabbit mAb and Ubiquitin Antibody #3933 (lower).
Western blot analysis of seven distinct recombinant polyubiquitin chains (300 ng each) using K63-linkage Specific Polyubiquitin (D7A11) Rabbit mAb (upper) and Ubiquitin Antibody #3933 (lower).
Western blot analysis comparing the titration of recombinant monoubiquitin, K48-linked polyubiquitin and K63-linked polyubiquitin using K63-linkage Specific Polyubiquitin (D7A11) Rabbit mAb (upper) and Ubiquitin Antibody #3933 (lower).
Western blot analysis of various cell lines using K63-linkage Specific Polyubiquitin (D7A11) Rabbit mAb.
Western blot analysis of extracts from HeLa cells using K63-linkage Specific Polyubiquitin (D7A11) Rabbit mAb, untreated or following antibody pre-incubation with either K63 ubiquitinylated branched peptide to block the signal or a linear peptide surrounding K63 of ubiquitin that cannot block the signal.
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).
Substrate proteins are linked to ubiquitin using seven distinct ubiquitin lysine residues (Lys6, Lys11, Lys27, Lys29, Lys33, Lys48, and Lys63). Formation of a polyubiquitin chain occurs when a lysine residue of ubiquitin is linked to the carboxy-terminal glycine of another ubiquitin. Proteins polyubiquitinated at specific lysine residues display a tendency to be targeted for different processes (8). K63-linked polyubiquitin chains exert nonproteolytic functions in vivo, such as protein trafficking, kinase/phosphatase activation, and DNA damage control, all of which might be important in regulation of cancer survival and development (9,10).
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- 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.
- Komander, D. (2009) Biochem Soc Trans 37, 937-53.
- Chen, Z.J. and Sun, L.J. (2009) Mol Cell 33, 275-86.
- Yang, W.L. et al. (2010) Oncogene 29, 4493-503.
- Tigno-Aranjuez, J.T. et al. (2013) Mol Cell Biol 33, 146-58. Applications: Western Blotting.
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