Anti-CDC27 phospho T244 Antibody

Human CDC27 (also called Cell division cycle protein 27 homolog, CDC27Hs and H-NUC) shares strong similarity with Saccharomyces cerevisiae protein Cdc27. This protein is a component of anaphase-promoting complex (APC), which is composed of eight protein subunits and highly conserved in eukaryotic cells. APC catalyzes the formation of a cyclin B-ubiquitin conjugate that is responsible for the ubiquitin-mediated proteolysis of B-type cyclins. This protein and 3 other members of the APC complex contain the TPR (tetratricopeptide repeat), a protein domain important for protein-protein interaction. This protein was shown to interact with mitotic checkpoint proteins including Mad2, p55CDC and BUBR1, and thus may be involved in controlling the timing of mitosis.

This affinity purified antibody was prepared from whole rabbit serum produced by repeated immunizations with a synthetic peptide corresponding to an internal portion surrounding T244 of Human CDC27.

This affinity purified antibody is directed against the phosphorylated form of human CDC27 at the pT244 residue. 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. Reactivity occurs against human CDC27 pT244 protein and the antibody is specific for the phosphorylated form of the protein. Reactivity with non-phosphorylated human CDC27 is minimal by ELISA. The antibody does not cross-react with CDC27 phosphorylated at other sites. A BLAST analysis was used to suggest reactivity with this protein from human, rat, dog, bovine and chimpanzee based on 100% homology for the immunogen sequence. Cross-reactivity with CDC27 protein from mouse, rat and chicken may occur as sequence homology varies by one amino acid residue in this sequence (90% homology) . Cross-reactivity with CDC27 homologues from other sources has not been determined.

Store vial at -20°C prior to opening. Aliquot contents and freeze at -20°C or below for extended storage. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4°C as an undiluted liquid. Dilute only prior to immediate use. Expiration date is one (1) year from date of opening.

This affinity purified antibody has been tested for use in ELISA and by western blot. Specific conditions for reactivity should be optimized by the end user. Expect a band approximately 92 kDa in size corresponding to CDC27 by western blotting in the appropriate cell lysate or extract. Less than 1% reactivity is observed against the non-phosphorylated form of the immunizing peptide. This antibody is phospho specific for pT244 of CDC27.