The production of affinity column purified (specific) anti-actin is described. With the immunization scheme employed, all rabbits produced precipitating antibodies over several months, so that 30 mg specific anti-actin per rabbit could be isolated in 6 months. The antibodies against native and detergent denatured smooth muscle actin are characterized by immunodiffusion tests, staining of the I-band of isolated myofibrils and stress fibers in tissue culture cells, using indirect immunofluorescence.