ELISA, Immunoprecipitation, Western Blot. 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 91 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 pS426 of CDC27.
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Description
This affinity purified antibody is directed against the phosphorylated form of human CDC27 at the pS426 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 pS426 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, chicken and chimpanzee based on 100% homology for the immunogen sequence. Cross reactivity with CDC27 protein from mouse may occur as sequence homology varies by one amino acid residue in this sequence (89% homology). Cross reactivity with CDC27 homologues from other sources has not been determined.
Immunogen
This affinity purified antibody was prepared from whole rabbit serum produced by repeated immunizations with a synthetic peptide corresponding aa 422-430 of Human CDC27.
Choi LJ, Jenikova G, Hanson E, Spehlmann ME, Boehling NS, Kirstein SL, Bundey RA, Smith JR, Insel PA, Eckmann L. (2010) Coordinate down-regulation of adenylyl cyclase isoforms and the stimulatory G protein (G(s)) in intestinal epithelial cell differentiation. J Biol Chem. 2010 Apr 23;285(17):12504-11. doi: 10.1074/jbc.M109.059741. Epub 2010 Feb 15.