SIMULTANEOUS DOUBLE IMMUNOENZYMATIC LABELING - A NEW PROCEDURE FOR THE HISTOPATHOLOGIC ROUTINE

We developed a new procedure for simultaneous immunohistoenzymatic dou ble labeling that uses two mouse monoclonal antibodies of different is otypes or two antibodies from different species. The technique is a co mbination of the alkaline phosphatase-antialkaline phosphatase, avidin -biotin, and digoxigenin methods and uses horseradish peroxidase and a lkaline-phosphatase detection systems. This standardized procedure can be carried out easily in the routine of a pathology laboratory and pe rformed in parallel with single-antigen immunostaining, without the ne ed for extra material or additional time for the laboratory technician s. This method can be applied to the detection of antigens localized i n distinct cells or subcellular compartments and to the covisualizatio n of different components from the same subcellular compartment.