Product Pathways - Protein Folding/Stability

NEDD8 (19E3) Rabbit mAb #2754

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Applications	Reactivity	Sensitivity	MW (kDa)	Source	Isotype
W IP IHC-P IF-IC F	H M R Mk (B) (X) (Z)	Endogenous	9	Rabbit	IgG

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IHC-P=Immunohistochemistry (Paraffin)  IF-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey  B=Bovine  X=Xenopus  Z=Zebra Fish
Species enclosed in parentheses are predicted to react based on 100% sequence homology. Species cross-reactivity is determined by Western blot.

Specificity / Sensitivity

NEDD8 (19E3) Rabbit mAb detects endogenous levels of both free and conjugated NEDD8 protein. The antibody does not cross-react with other ubiquitin family members, including ubiquitin, SUMO-1, SUMO-2, SUMO-3 and ISG15.

Source / Purification

Monoclonal antibody is produced by immunizing rabbits with a synthetic peptide (KLH-coupled) corresponding to amino acids at the amino terminus of human NEDD8 protein.

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).

1. Chiba, T. and Tanaka, K. (2004) Curr. Protein Pept. Sci. 5, 177-184.
2. Schwartz, D.C. and Hochstrasser, M. (2003) Trends Biochem. Sci. 28, 321-328.
3. Wada, H. et al. (1998) Biochem. Biophys. Res. Commun. 251, 688-692.
4. Hemelaar, J. et al. (2004) Mol. Cell Biol. 24, 84-95.
5. Osaka, F. et al. (1998) Genes Dev. 12, 2263-2268.
6. Kamura, T. et al. (1999) Genes Dev. 13, 2928-2933.
7. Gong, L. et al. (2000) J. Biol. Chem. 275, 14212-14216.
8. Mendoza, H.M. et al. (2003) J. Biol. Chem. 278, 25637-25643.
9. Lyapina, S. et al. (2001) Science 292, 1382-1385.
10. Kawakami, T. et al. (2001) EMBO J. 20, 4003-4012.
11. Stickle, N.H. et al. (2004) Mol. Cell Biol. 24, 3251-3261.
12. Xirodimas, D.P. et al. (2004) Cell 118, 83-97.

Application References

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Rabbit Monoclonals Produced Using Epitomics® Technology, U.S. Patent No. 5,675,063.

This product is for in vitro research use only and is not intended for use in humans or animals. This product is not intended for use as therapeutic or in diagnostic procedures.