ELISA, Immunohistochemistry, Western Blot. This affinity purified antibody has been tested for use in ELISA, immunohistochemistry and by western blot. Specific conditions for reactivity should be optimized by the end user. Expect a band approximately 47 kDa in size corresponding to phosphorylated E2F-1 by western blotting in the appropriate cell lysate or extract. Less than 0.5% reactivity is observed against the non-phosphorylated form of the immunizing peptide. This antibody is phospho specific for pS364 of E2F-1.
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Description
This affinity purified antibody is directed against the phosphorylated form of human E2F-1 at the pS364 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 E2F-1 pS364 protein and the antibody is specific for the phosphorylated form of the protein. Reactivity with non-phosphorylated human E2F-1 is minimal by ELISA. The antibody does not cross-react with E2F-1 phosphorylated at other sites. A BLAST analysis was used to suggest reactivity with this protein from human and chimpanzee based on 100% homology for the immunogen sequence. Cross reactivity with E2F-1 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 to amino acids 360-369 of Human E2F-1.