Quantitation of peroxidase-antibody binding to membrane fragments using column chromatography.

A method which can be used to measure the amount of peroxidase antibody which is specifically bound to placental alkaline phosphatase in membrane fragments is described. The technique uses Sepharose 4B chromatography to separate membrane fragments containing bound peroxidase-antibody from unbound peroxidase-antibody. Specificity is demonstrated by nonbinding to rabbit placental membranes, by a strict correlation between membrane-associated phosphatase and bound peroxidase-antibody, and by preventing binding using pure enzyme. The general utility of this method for membrane antigens is discussed.