ab12281 is specific for the phosphorylated form of the Cdc27 (phospho T244). Less than 1% reactivity is observed against the non-phosphorylated form of the immunizing peptide. The antibody does not cross-react with Cdc27 phosphorylated at other sites.
The product was affinity purified from monospecific antiserum by immunoaffinity purification. Antiserum was first purified against the phosphorylated form of the immunizing peptide. The resultant affinity purified antibody was then cross adsorbed against the non-phosphorylated form of the immunizing peptide.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
ELISA: 1/3000 - 1/15000. Reactivity with non-phosphorylated human Cdc27 is minimal by ELISA.
IF: Use at an assay dependent dilution.
WB: 1/300 - 1/2000. Detects a band of approximately 92 kDa.
ICC/IF: Use at an assay dependent dilution (from PubMed: 17472438).
Not tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
Component of the anaphase promoting complex/cyclosome (APC/C), a cell cycle-regulated E3 ubiquitin ligase that controls progression through mitosis and the G1 phase of the cell cycle. The APC/C complex acts by mediating ubiquitination and subsequent degradation of target proteins: it mainly mediates the formation of 'Lys-11'-linked polyubiquitin chains and, to a lower extent, the formation of 'Lys-48'- and 'Lys-63'-linked polyubiquitin chains.
Protein modification; protein ubiquitination.
Belongs to the APC3/CDC27 family. Contains 9 TPR repeats.
Phosphorylated. Phosphorylation on Ser-426 and Thr-446 occurs specifically during mitosis.
Skoufias DA et al. Mitosis persists in the absence of Cdk1 activity when proteolysis or protein phosphatase activity is suppressed. J Cell Biol179:671-85 (2007).
Read more (PubMed: 18025303) »
Lindqvist A et al. Cyclin B1-Cdk1 activation continues after centrosome separation to control mitotic progression. PLoS Biol5:e123 (2007).
Read more (PubMed: 17472438) »