The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 2.5 µg/ml. Detects a band of approximately 20 kDa (predicted molecular weight: 21.5 kDa).Can be blocked with Human UBE2C peptide (ab22967).
Use at an assay dependent dilution. (See Abreview)
1/200. (See Abreview)
Accepts ubiquitin from the E1 complex and catalyzes its covalent attachment to other proteins. In vitro catalyzes 'Lys-11'- and 'Lys-48'-linked polyubiquitination. Acts as an essential factor of the anaphase promoting complex/cyclosome (APC/C), a cell cycle-regulated ubiquitin ligase that controls progression through mitosis. Acts by initiating 'Lys-11'-linked polyubiquitin chains on APC/C substrates, leading to the degradation of APC/C substrates by the proteasome and promoting mitotic exit.
Protein modification; protein ubiquitination.
Belongs to the ubiquitin-conjugating enzyme family.
Autoubiquitinated by the APC/C complex, leading to its degradation by the proteasome. Its degradation plays a central role in APC/C regulation, allowing cyclin-A accumulation before S phase entry. APC/C substrates inhibit the autoubiquitination of UBE2C/UBCH10 but not its E2 function, hence APC/C remaining active until its substrates have been destroyed.
ab3935 staining approximately 20 kDa human UBE2 after immmunoprecipitation from HeLa whole cell lysate (106 cells/ml) using 40µl of ab3935 saturated protein G dynabeads, by Western blot (ECL):
Lane 1 = IP from Noc arrested HeLa cell extract (beads only). Lane 2 = IP from Noc arrested HeLa cell extract (goat whole serum coupled to beads). Lane 3 = IP from Noc arrested HeLa cell extract (ab3935 coupled to beads) Lane 4 = Noc arrested HeLa whole cell lysate.
For a more detailed description, please click the reviews tab at the top of the page and see the review dated 25/02/03.
ab3935 at a 1/200 dilution staining human UBE2C in paraformaldehyde fixed HeLa cells by Immunocytochemistry. In the upper panel G2 phase cells can be seen (G1 cells stained only faintly). The lower panel shows a mitotic (metaphase) cell. Diffuse cytoplamic staining with some microtubule localisation can be observed in mitotic cells.