The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/1000 - 1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
1/2000 - 1/10000. Detects a band of approximately 18 kDa (predicted molecular weight: 18 kDa).
Use at 2-5 µg/mg of lysate.
Efficiency in IP is low.
Accepts ubiquitin from the E1 complex and catalyzes its covalent attachment to other proteins. In vitro catalyzes 'Lys-11'-linked polyubiquitination. Involved in the selective degradation of short-lived and abnormal proteins. Down-regulated during the S-phase it is involved in progression through the cell cycle. Regulates nuclear hormone receptors transcriptional activity. May play a role in myelopoiesis.
Ubiquitous, with highest expression in testis.
Protein modification; protein ubiquitination.
Belongs to the ubiquitin-conjugating enzyme family.
Ubiquitinated. The alteration of UBE2L3 protein levels during the S-phase of the cell cycle is due to ubiquitin-dependent proteasomal degradation.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human prostate carcinoma tissue labelling Ube2L3 with ab72545 at 1/1000 (1µg/ml). Detection: DAB.
Western blot - Ube2L3 antibody (ab72545)
All lanes : Anti-Ube2L3 antibody (ab72545) at 0.1 µg/ml
Lane 1 : HeLa whole cell lysate at 50 µg Lane 2 : HeLa whole cell lysate at 15 µg Lane 3 : HeLa whole cell lysate at 5 µg Lane 4 : 293T whole cell lysate at 50 µg Lane 5 : NIH3T3 whole cell lysate at 50 µg
Predicted band size : 18 kDa Observed band size : 18 kDa
Immunoprecipitation - Ube2L3 antibody (ab72545)
Detection of Human Ube2L3 by Immunoprecipitation from Whole cell lysate from HeLa cells (1 mg for IP, 20% of IP loaded) using ab72545 at 3 µg/mg lysate for IP (Lane 1) and at 1 µg/ml for subsequent WB detection. Lane 2 represents control IgG IP.