WB, IP, IHC-P
H M R Mk
Endogenous
9
Rabbit
#Q15843
4738
Product Information
Product Usage Information
Application | Dilution |
---|---|
Western Blotting | 1:1000 |
Immunoprecipitation | 1:25 |
Immunohistochemistry (Paraffin) | 1:200 |
Storage
Specificity / Sensitivity
Species Reactivity:
Human, Mouse, Rat, Monkey
Species predicted to react based on 100% sequence homology
The antigen sequence used to produce this antibody shares
100% sequence homology with the species listed here, but
reactivity has not been tested or confirmed to work by CST.
Use of this product with these species is not covered under
our
Product Performance Guarantee.
Xenopus, Zebrafish, Bovine
Source / Purification
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to amino acids at the amino-terminus of human NEDD8 protein. Antibodies are purified by protein A and peptide affinity chromatography.
Background
Neural precursor cell-expressed developmentally downregulated protein 8 (NEDD8), also known as Rub1 (related to ubiquitin 1) in plants and yeast, is a member of the ubiquitin-like protein family (1,2). The covalent attachment of NEDD8 to target proteins, termed neddylation, is a reversible, multi-step process analogous to ubiquitination. NEDD8 is first synthesized in a precursor form with a carboxy-terminal extension peptide that is removed by either the UCH-L3 or NEDP1/DEN1 hydrolase protein to yield a mature NEDD8 protein (3,4). Mature NEDD8 is then covalently linked to target proteins via the carboxy-terminal glycine residue in a reaction catalyzed by the APP-BP1/Uba3 heterodimer complex and Ubc12 as the E1- and E2-like enzymes, respectively (5). An E3 ligase protein, Roc1/Rbx1, is also required for neddylation of the cullin proteins (6). Protein de-neddylation is catalyzed by a number of enzymes in the cell, including a "ubiquitin-specific" protease USP21, the NEDP1/DEN1 hydrolase and the COP9/signalosome (CSN) (7,8,9). In contrast to the ubiquitin pathway, the NEDD8 modification system acts on only a few substrates and does not appear to target proteins for degradation. Neddylation of cullin proteins activates the SCF (Skp1-Cullin-F-box) E3 ubiquitin ligase complex by promoting complex formation and enhancing the recruitment of the E2-ubiquitin intermediate (10). While NEDD8 modification of VHL is not required for ubiquitination of HIF1-α, it is required for fibronectin matrix assembly (11). Mdm2-dependent neddylation of p53 inhibits its transcriptional activity (12).
- Chiba, T. and Tanaka, K. (2004) Curr. Protein Pept. Sci. 5, 177-184.
- Schwartz, D.C. and Hochstrasser, M. (2003) Trends Biochem. Sci. 28, 321-328.
- Wada, H. et al. (1998) Biochem. Biophys. Res. Commun. 251, 688-692.
- Hemelaar, J. et al. (2004) Mol. Cell Biol. 24, 84-95.
- Osaka, F. et al. (1998) Genes Dev. 12, 2263-2268.
- Kamura, T. et al. (1999) Genes Dev. 13, 2928-2933.
- Gong, L. et al. (2000) J. Biol. Chem. 275, 14212-14216.
- Mendoza, H.M. et al. (2003) J. Biol. Chem. 278, 25637-25643.
- Lyapina, S. et al. (2001) Science 292, 1382-1385.
- Kawakami, T. et al. (2001) EMBO J. 20, 4003-4012.
- Stickle, N.H. et al. (2004) Mol. Cell Biol. 24, 3251-3261.
- Xirodimas, D.P. et al. (2004) Cell 118, 83-97.
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 BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
Applications Key
WB: Western Blotting IP: Immunoprecipitation IHC-P: Immunohistochemistry (Paraffin)
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 All: all species expected
Trademarks and Patents
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