The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 1 µg/ml. Predicted molecular weight: 31 kDa. Good results were obtained when blocked with 5% non-fat dry milk in 0.05% PBS-T.
FunctionE3 ubiquitin-protein ligase that mediates ubiquitination of TFRC, CD86, FAS and MHC class II proteins, such as HLA-DR alpha and beta, and promotes their subsequent endocytosis and sorting to lysosomes via multivesicular bodies. By constitutively ubiquitinating MHC class II proteins in immature dendritic cells, downregulates their cell surface localization thus sequestering them in the intracellular endosomal system.
Tissue specificityExpressed in antigen presenting cells, APCs, located in lymph nodes and spleen. Also expressed in lung. Expression is high in follicular B cells, moderate in dendritic cells and low in splenic T cells.
PathwayProtein modification; protein ubiquitination.
Developmental stageDuring maturation of dendritic cells, expression is down-regulated and stabilizes MHC class II proteins accumulate at the plasma membrane.
DomainThe RING-CH-type zinc finger domain is required for E3 ligase activity.
Post-translational modificationsHas a short half-life. Instability/short half-life permits rapid changes that allow efficient induction of antigen presentation once antigen presenting cells, APCs, receive maturation signals. Small changes in protein levels significantly alter the cell surface display of MHC class II proteins.
Cellular localizationGolgi apparatus > trans-Golgi network membrane. Lysosome membrane. Cytoplasmic vesicle membrane. Late endosome membrane. Early endosome membrane. Cell membrane.