Development and characterization of cholinephosphotransferase antibody

In the present study, we generated antibodies in rabbits against two synthe tic peptides, one based on peptide sequence from yeast CPT cDNA (position 8 6 to 98 of the amino acid sequence) and the other from our guinea pig CPT c DNA (it corresponds to amino acid positions 119 to 130 according to yeast C PT gene). The antibody titers were measured by both dot blot analysis and E LISA using Keyhole limpets hemocyanin coupled CPT peptides. The CPT antibod y recognized a single band by Western blot analysis of proteins from guinea pig liver mitochondria and microsomes. The molecular weight of the protein recognized by Western blot analysis is close to the predicted molecular we ight (46 kDa) of yeast CPT. Further analysis revealed that the antibody inh ibited CPT activity in both subcellular fractions in a dose dependent manne r, thus confirming the specificity of the antibody against both subcellular CPT. (C) 2001 Academic Press.