The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use at an assay dependent concentration.
ELISA titre using peptide based assay, 1/:2500.
Use a concentration of 1 µg/ml. Predicted molecular weight: 61 kDa. Good results were obtained when blocked with 5% non-fat dry milk in 0.05% PBS-T.
FunctionE3 ubiquitin-protein ligase. Mediates monoubiquitination at multiple sites of TSG101 in the presence of UBE2D1, but not of UBE2G1, nor UBE2H. Plays a role in the regulation of endosome-to-lysosome trafficking. Impairs MC1R- and MC4R-signaling by competing with GNAS-binding to MCRs and inhibiting agonist-induced cAMP production. Does not inhibit ADRB2-signaling. Does not promote MC1R ubiquitination.
PathwayProtein modification; protein ubiquitination.
DomainThe RING finger is required for ubiquitin ligase activity.
Post-translational modificationsAutoubiquitinated in vitro.
Cellular localizationCytoplasm > cytosol. Cell membrane; Early endosome. The endosomal localization is dependent on the interaction with TSG101 and Cytoplasm > cytosol. Nucleus. Translocation from the cytosol to the nucleus is seen only in the presence of MC1R and MC4R, but not TBXA2R. Excluded from nucleoli.