ELISA, Western Blot. This affinity purified antibody has been tested for use in ELISA and western blotting using M1 cells which express endogenous wild-type c-Myb protein and in COS7 cells transfected with c-Myb. Specific conditions for reactivity and detection of phosphorylated c-Myb should be optimized by the end user. Expect a band approximately ~75 kDa in size corresponding to phosphorylated c-Myb by western blotting in the appropriate cell lysate or extract.
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Description
This affinity-purified antibody is directed against the phosphorylated form of mouse c-Myb protein at the pT486 residue. 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 mouse c-Myb pT486 protein and the antibody is specific for the phosphorylated form of the protein. This antibody reacts with endogenous c-Myb protein and also SUMOylated overexpressed c-Myb protein in transfected Cos7 cells. Reactivity with non- phosphorylated mouse c-Myb is minimal by ELISA and western blot. A BLAST analysis was used to suggest reactivity with c-Myb from mouse based on a 100% homology with the immunizing sequence. Expect reactivity with c-Myb from rat, pig, human, horse, platypus, opossum, orangutan, spider monkey, bonobo, gorilla, and chimpanzee; with a v-myb myeloblastosis viral oncogene homolog, all isoforms (also avian version) from human; and with a protooncogene c-myb, and a v-myb myeloblastosis viral oncogene homolog (avian), both from cattle, all based on a 93% homology with the immunizing sequence. Cross-reactivity with c-Myb 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 an internal region of mouse c-Myb protein surrounding amino acid residue 486.