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Product Pathways - Protein Stability

NEDD8 (19E3) Rabbit mAb #2754

Item# Description List Price Web Price Qty
2754S NEDD8 (19E3) Rabbit mAb - 100 µl $372.00
$334.80
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2754T NEDD8 (19E3) Rabbit mAb - 20 µl $159.00
$143.10
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VIEW COMPANION PRODUCTS HIDE COMPANION PRODUCTS
Application Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W Human, Mouse, Rat, Monkey Endogenous 9 Rabbit IgG
IP
IHC-P
F

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting, IP=Immunoprecipitation, IHC-P=Immunohistochemistry (Paraffin), F=Flow Cytometry

Homology

Species predicted to react based on 100% sequence homology: Xenopus, Zebrafish, Bovine.

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 animals with a synthetic peptide corresponding to amino acids at the amino terminus of human NEDD8 protein.

Western Blotting

Western Blotting

Western blot analysis of ubiquitin, NEDD8, ISG15 and SUMO-2/3 recombinant proteins (5 ng each), using NEDD8 (19E3) Rabbit mAb, Ubiquitin (P4D1) Mouse mAb #3936, ISG15 Antibody #2743 and SUMO-2/3 Antibody #4974.

Western Blotting

Western Blotting

Western blot analysis of lysates from HeLa, RAW, C6 and COS cells, using NEDD8 (19E3) Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human colon carcinoma, using NEDD8 (19E3) Rabbit mAb.


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human ovarian carcinoma, using NEDD8 (19E3) Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma, using NEDD8 (19E3) Rabbit mAb, preincubated with control peptide (left) or Nedd8 Blocking Peptide #1048 (right).

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of untreated HeLa cells using NEDD8 (19E3) Rabbit mAb (blue) compared to a nonspecific negative control antibody (red).


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

  • Hjerpe, R. et al. (2012) Biochem J 441, 927-36. Applications: Western Blotting.

Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!


 

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