Production and characterization of monoclonal antibody against 10S DNA polymerase alpha from calf thymus.
One hybridoma cell line that produces an antibody directed against 10S DNA polymerase alpha 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 alpha and 6.5S DNA polymerase alpha-2 from calf thymus, but not 6.5S DNA polymerase alpha-1. The antibody precipitated both polypeptides of 140-150,000 and 46-50,000 dalton of 10S DNA polymerase alpha. The antibody also recognized the DNA polymerase alpha purified from human cells, but did pig DNA polymerase alpha only partially. The antibody did not crossreact with rat DNA polymerase alpha, calf DNA polymerase beta, 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.
D000818:Animals; D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D002417:Cattle; D002459:Cell Fusion; D002460:Cell Line; D003429:Cross Reactions; D004257:DNA Polymerase II / Q000276:immunology*; D004259:DNA-Directed DNA Polymerase / Q000276:immunology*; D006825:Hybridomas / Q000276:immunology; D008214:Lymphocytes / Q000276:immunology; D051379:Mice; D008815:Mice, Inbred Strains; D008970:Molecular Weight; D010954:Plasmacytoma / Q000276:immunology; D013950:Thymus Gland / Q000201:enzymology*
D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D004257:DNA Polymerase II; D004259:DNA-Directed DNA Polymerase