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 C-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-alpha, it is required for fibronectin matrix assembly (11).
Mdm2-dependent neddylation of p53 inhibits its transcriptional activity (12).
Applications:Suitable for use in Western Blot, Immunohistochemistry, Immunofluorescence/Immunocytochemistry and ELISA.
Other applications not tested.
Recommended Dilution:Western Blot: 1:500-1:2000, incubate membrane with diluted primary antibody in TBS, 5% milk, 0.1% Tween 20 at 4°C with gentle shaking, overnight
Immunohistochemistry: 1:50-1:200Immunofluorescence (IF-IC): 1:100-1:500ELISA (peptide): 1:20,000-1:40,000Optimal dilutions to be determined by the researcher.
Storage and Stability:May be stored at 4°C for short-term only.
Aliquot to avoid repeated freezing and thawing.
Store at -20°C.
Aliquots are stable for 12 months after receipt.
For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
仕様
Size：200ul
Host：rabbit
Source Antibody：human
Grade：Affinity Purified
Purity：Purified by immunoaffinity chromatography from serum.
Form：Supplied as a liquid in PBS, pH 7.4, 0.02% sodium azide, 50% glycerol.
Specificity：Recognizes endogenous levels of total human NEDD8. Species Crossreactivity: Mouse and rat
Isotype：IgG
Calc Applications Abbrev：E IC IF IHC WB
Calc Crossreactivity：Hu Mo Rt
Immunogen：Synthetic peptide corresponding to human NEDD8.
