The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Biological activitySpecific Activity: 0.0027 pmol/min/µg.
Assay conditions: 50 µl reaction mix (4mM ATP, 5mM MgCl2, 1mM DTT, E1 (0.5 µg), E2 (2.8 µg), Biotin-Ubiquitin (0.5 µg) and NEDD4 (0-2 µg) in 50mM Tris pH 7.4) incubate for 30min at 37°C, then add to streptavidin coated plates and incubate for 30min at 37°C. Add antibody against NEDD4, incubate 1h. Finally, incubate 1h with the Europeum-labeled secondary antibody. Time resolved fluorescence measured at room temperature (Lambda exc=330 nm, Lambda em=620 nm).
% . Purity is >60% by SDS-PAGE.
Concentration information loading...
Preparation and Storage
Stability and Storage
Shipped on Dry Ice. Store at -80°C.
pH: 8.00 Constituents: 20% Glycerol, 0.58% Sodium chloride, 0.395% Tris HCl, 0.05% Tween, 0.0462% DTT Note: 100 µg/ml FLAG peptide
This product is an active protein and may elicit a biological response in vivo, handle with caution.
Cell proliferation-inducing gene 53 protein
E3 ubiquitin protein ligase Nedd4
E3 ubiquitin-protein ligase NEDD4
HECT-type E3 ubiquitin transferase NEDD4
Neural precursor cell expressed developmentally down regulated 4
Neural precursor cell expressed developmentally down-regulated protein 4
FunctionE3 ubiquitin-protein ligase which accepts ubiquitin from an E2 ubiquitin-conjugating enzyme in the form of a thioester and then directly transfers the ubiquitin to targeted substrates. Involved in the pathway leading to the degradation of VEGFR-2/KDFR, independently of its ubiquitin-ligase activity. Monoubiquitinates IGF1R at multiple sites, thus leading to receptor internalization and degradation in lysosomes. According to PubMed:18562292 the direct link between NEDD4 and PTEN regulation through polyubiquitination described in PubMed:17218260 is questionable. Involved in ubiquitination of ERBB4 intracellular domain E4ICD. Involved in the budding of many viruses. Part of a signaling complex composed of NEDD4, RAP2A and TNIK which regulates neuronal dendrite extension and arborization during development. Ubiquitinates TNK2 and regulates EGF-induced degradation of EGFR and TNF2.
PathwayProtein modification; protein ubiquitination.
Cellular localizationCytoplasm. Cell membrane. Recruited to the plasma membrane by GRB10. Once complexed with GRB10 and IGF1R, follows IGF1R internalization, remaining associated with early endosomes. Uncouples from IGF1R-containing endosomes before the sorting of the receptor to the lysosomal compartment (By similarity). May be recruited to exosomes by NDFIP1.