Production of specific antibodies to contractile proteins and their use in immunofluorescence microscopy

Summary The preparation of highly purified myosin from surgical specimen of human uterine muscle is described. Antibodies were raised in rabbits against this immunogen. In immunodiffusion, they react with uterine and chicken gizzard muscle myosin, no reaction is observed between uterine myosin and the anti-chicken-gizzard- myosin. In immunofluorescence, antiuterine-myosin stains smooth muscle in the contractile and modulated state and non-muscle cells such as fibroblasts, platelets and endothelium of various species. Thus, these antibodies contrast anti-gizzard-myosin, which has previously been shown to be specific for contractile state muscle cells. We therefore conclude that the uterine myosin preparation consists of two immunogens, the one being associated with cell contractility and the other, termed cytoplasmic myosin, with motility and mitosis. The latter is indistinguishable from the myosin present in non-muscle cells and can be absorbed specifically with actomyosin from blood platelets.Abbreviations ATP Adenosine triphosphate - DNAse I Deoxyribonuclease I - DTE Dithioerythritol - SDS Sodiumdodecylsulfate - PAGE Polyacrylamide electrophoresis