The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Specific 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.
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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
E3 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.
The WW domains mediate interaction with LITAF, RNF11, WBP1, WBP2, TMEPAI, NDFIP1 and PRRG2.
Cytoplasm. 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.