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.
E3 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.
Expressed 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.
Protein modification; protein ubiquitination.
Contains 1 RING-CH-type zinc finger.
During maturation of dendritic cells, expression is down-regulated and stabilizes MHC class II proteins accumulate at the plasma membrane.
The RING-CH-type zinc finger domain is required for E3 ligase activity.
Has 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.
Golgi apparatus > trans-Golgi network membrane. Lysosome membrane. Cytoplasmic vesicle membrane. Late endosome membrane. Early endosome membrane. Cell membrane.