Product Pathways - Protein Stability
NEDD8 Antibody #2745
|W IP IHC-P F||H M R Mk (X) (Z) (B)||Endogenous||9||Rabbit|
Reactivity Key: H=Human M=Mouse R=Rat Mk=Monkey X=Xenopus Z=Zebrafish B=Bovine
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.
Specificity / Sensitivity
This antibody detects endogenous levels of both free and conjugated NEDD8 protein. The antibody does not cross-react with other ubiquitin family members, including ubiquitin, SUMO1, SUMO2, SUMO3 and ISG15.
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.
Western blot analysis of Ubiquitin, NEDD8, ISG15 and SUMO-2/3 recombinant proteins (5 ng each), using NEDD8, Ubiquitin, ISG15 and SUMO-2/3 Antibodies.
Western blot analysis of lysates from HeLa, Raji, RAW, C6 and COS cells, using NEDD8 antibody.
Immunohistochemical analysis of paraffin-embedded human colon carcinoma, showing cytoplasmic and nuclear localization, using NEDD8 Antibody.
Immunohistochemical analysis of paraffin-embedded human breast carcinoma, using NEDD8 Antibody in the presence of control peptide (left) or Nedd8 Blocking Peptide #1048 (right).
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.
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For Research Use Only. Not For Use In Diagnostic Procedures.