Cell Signaling Technology

Product Pathways - Protein Stability

NEDD4 Antibody #2740

Applications Reactivity Sensitivity MW (kDa) Source
W IF-IC F H M R Mk Endogenous 115 kDa Rabbit

Applications Key:  W=Western Blotting  IF-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

This antibody detects endogenous levels of total NEDD4 protein. The antibody may also recognize other NEDD4-like proteins, including NEDD4L (NEDD4-2).

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to human NEDD4 protein. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of lysates from NIH/3T3, C6, H-4-II-E, U87, A549 and COS cells, using NEDD4 antibody.

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of NIH/3T3 cells, using NEDD4 Antibody (blue) compared to a nonspecific negative control antibody (red).

IF-IC

IF-IC

Immunofluorescent analysis of paraformaldehyde-fixed NIH/3T3 cells, using NEDD4 antibody.


IF-IC

IF-IC

Confocal immunofluorescent analysis of untreated C6 cells using NEDD4 Antibody (green). Blue pseudocolor = DRAQ5® #4084 (DNA fluorescent dye).

Background

Neural precursor expressed, developmentally down-regulated protein 4 (NEDD4) was originally identified as a gene that is highly expressed in the early mouse embryonic central nervous system (1). Subsequently, a family of NEDD4-like proteins have been defined that includes seven members in humans (2). NEDD4 and NEDD4-like (NEDD4L) proteins contain multiple functional domains including a calcium-dependent phospholipid and membrane binding domain (C2 domain), two to four protein binding domains (WW domains), and an E3 ubiquitin-protein ligase domain (HECT domain). NEDD4 and NEDD4L have been shown to downregulate both neuronal voltage-gated Na+ channels (NaVs) and epithelial Na+ channels (ENaCs) in response to increased intracellular Na+ concentrations (3,4). The WW domains of NEDD4 bind to PY motifs (amino acid sequence PPXY) found in multiple NaV and ENaC proteins; ubiquitination of these proteins is mediated by the HECT domain of NEDD4 and results in their internalization and removal from the plasma membrane. Research studies have shown that mutation of the PY motifs in ENaC proteins is associated with Liddle's syndrome, an autosomal dominant form of hypertension (5). In addition to targeting sodium channels, NEDD4L has also been shown to negatively regulate TGF-β signaling by targeting Smad2 for degradation (6). Mouse and human NEDD4 are rapidly cleaved by caspase proteins during apoptosis, although the significance of this cleavage is not clear (7).

  1. Kumar, S. et al. (1992) Biochem Biophys Res Commun 185, 1155-61.
  2. Harvey, K.F. and Kumar, S. (1999) Trends Cell Biol 9, 166-9.
  3. Dinudom, A. et al. (1998) Proc Natl Acad Sci USA 95, 7169-73.
  4. Goulet, C.C. et al. (1998) J Biol Chem 273, 30012-7.
  5. Staub, O. et al. (1996) EMBO J 15, 2371-80.
  6. Kuratomi, G. et al. (2005) Biochem J 386, 461-70.
  7. Harvey, K.F. et al. (1998) J Biol Chem 273, 13524-30.

Application References

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

Companion Products

For Research Use Only. Not For Use In Diagnostic Procedures.

Products