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Immunohistochemical analysis of paraffin-embedded human breast carcinoma using NEDD8 (19E3) Rabbit mAb #2754 in the presence of control peptide (left) or NEDD8 Blocking Peptide (right).

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Product Description

This peptide is used to block NEDD8 (19E3) Rabbit mAb #2754 reactivity.

Quality Control

The quality of the peptide was evaluated by reversed-phase HPLC and by mass spectrometry. The peptide blocks NEDD8 (19E3) Rabbit mAb #2754 by dot blot.

Product Usage Information

Use as a blocking reagent to evaluate the specificity of antibody reactivity in dot blot protocols.

Storage: Supplied in 20 mM potassium phosphate (pH 7.0), 50 mM NaCl, 0.1 mM EDTA, 1 mg/ml BSA and 5% glycerol. 1% DMSO. Store at –20°C.

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, 321-8.

3.  Wada, H. et al. (1998) Biochem Biophys Res Commun 251, 688-92.

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, 2928-33.

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.

Entrez-Gene Id 4738
Swiss-Prot Acc. Q15843

For Research Use Only. Not For Use In Diagnostic Procedures.
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NEDD8 Blocking Peptide