CST, 2745S, NEDD8 Antibody

Polyclonal Antibody for studying NEDD8. Validated for Western Blotting,Immunoprecipitation,Immunohistochemistry (Paraffin). Highly specific and rigorously validated in-house, NEDD8 Antibody (CST #2745) is ready to ship. Product Usage Information Western Blotting: 1:1000 Immunoprecipitation: 1:25 Immunohistochemistry (Paraffin): 1:200 Storage Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA and 50% glycerol. Store at -20°C. Do not aliquot the antibody. Protocol Available protocols: Western Blotting, Immunoprecipitation, Immunohistochemistry (Paraffin) Specificity / Sensitivity NEDD8 Antibody detects endogenous levels of both free and conjugated NEDD8 protein. This antibody does not cross-react with other ubiquitin family members, including ubiquitin, SUMO1, SUMO2, SUMO3 and ISG15. Species Reactivity: Human, Mouse, Rat, Monkey 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). Alternate Names FLJ43224; MGC104393; MGC125896; MGC125897; NEDD-8; NEDD8; NEDD8 ubiquitin like modifier; Neddylin; Neural precursor cell expressed developmentally down-regulated protein 8; neural precursor cell expressed, developmentally down-regulated 8; Ubiquitin-like protein Nedd8 Specification REACTIVITY: H M R Mk SENSITIVITY: Endogenous MW (kDa): 9 SOURCE: Rabbit