Production and characterization of monoclonal antibody against 10S DNA polymerase a from calf thymus

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Abstract
One hybridoma cell line that produces an antibody directed against 10S DNA polymerase a purified from calf thymus was obtained. The monoclonality of the antibody was tested by sodium dodecyl sulfate polyacrylamide gel electrophoresis, isoelectrofocusing and antibody subclass determination. The antibody specifically recognized the 10S DNA polymerase α and 6.5S DNA polymerase α-2 from calf thymus, but not 6. 5S DNA polymerase α-1. The antibody precipitated both polypeptides of 140-150,000 and 46-50,000 dalton of 10S DNA polymerase α. The antibody also recognized the DNA polymerase α purified from human cells, but did pig DNA polymerase α only partially. The antibody did not crossresct with rat DNA polymerase α, calf DNA polymerase β, virus DNA polymerase and E. coli DNA polymerase I. This antibody will be a useful tool for studying the mechanism of DNA replication in eukaryotic cells.