antibody

Nedd8 (Rub1)

N0580-02A

100 ul

polyclonal

domestic rabbit

nonconjugated

Nedd8 (Rub1)

rabbit

Pab

Antibodies

100 ul

IgG

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- , it is required for fibronectin matrix assembly (11). Mdm2-dependent neddylation of p53 inhibits its transcriptional activity (12). Applications: Suitable for use in Immunofluorescence, Western Blot, Immunoprecipitation, Immunohistochemistry (paraffin), and Flow Cytometry. Other applications not tested. Recommended Dilution: Western Blot 1:1000 Immunoprecipitation 1:25 Immunofluorescence (IF-IC) 1:200 Flow Cytometry: 1:200 Optimal dilutions to be determined by the researcher. Storage and Stability: May be stored at 4°C for short-term only. For long-term storage, store at -20°C. Aliquots are stable for at least 12 months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.

Hu Mk Mo Rt

Synthetic peptide corresponding to amino acids at the N-terminus of human NEDD8 (KLH-coupled). Species sequence homology: bovine, Xenopus, and zebrafish

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. Species Crossreactivity: human, monkey, mouse and rat.

Purified by immunoaffinity chromatography

Supplied as a liquid in 10mM HEPES, pH 7.5, 150mM sodium chloride, 100ug/ml BSA, <0.02% sodium azide, 50% glycerol.

Not determined

Product Type: Pab Isotype: IgG Host: rabbit Source: human Concentration: Not determined Form: Supplied as a liquid in 10mM HEPES, pH 7.5, 150mM sodium chloride, 100ug/ml BSA, <0.02% sodium azide, 50% glycerol. Purity: Purified by immunoaffinity chromatography. Immunogen: Synthetic peptide corresponding to amino acids at the N-terminus of human NEDD8 (KLH-coupled). Species sequence homology: bovine, Xenopus, and zebrafish Specificity: 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. Species Crossreactivity: human, monkey, mouse and rat. Important Note: This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.

FC IC IF IP WB

-20°C

Background References: (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.

https://usbio-images.r.worldssl.net/prodimages/38/N0580-02A_1.jpg

Immunofluorescent analysis of paraformaldehyde-fixed HeLa cells, using N0580-02A.