The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 1 - 5 µg/ml. Detects a band of approximately 36.01 kDa (predicted molecular weight: 118 kDa). This antibody has only been tested in WB against the recombinant fragment used as immunogen. We have no data on the detection of endogenous protein
Use a concentration of 3 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Use a concentration of 10 µg/ml.
FunctionActivates ubiquitin by first adenylating its C-terminal glycine residue with ATP, and thereafter linking this residue to the side chain of a cysteine residue in E1, yielding an ubiquitin-E1 thioester and free AMP. Specific for ubiquitin, does not activate ubiquitin-like peptides. Differs from UBE1 in its specificity for substrate E2 charging. Does not charge cell cycle E2s, such as CDC34. Essential for embryonic development. Required for UBD/FAT10 conjugation. Isoform 2 may play a key role in ubiquitin system and may influence spermatogenesis and male fertility.
Tissue specificityWidely expressed. Isoform 2 is predominantly expressed in testis with higher expression in adult testis than in fetal testis.
PathwayProtein modification; protein ubiquitination.
Sequence similaritiesBelongs to the ubiquitin-activating E1 family.