Microwaving for double indirect immunofluorescence with primary antibodiesfrom the same species and for staining of mouse tissues with mouse monoclonal antibodies

Many methods have been devised for double immunocytochemical staining. We n ow describe that moderate microwaving does not elute antibodies, but preven ts their reactions with subsequently applied reagents. Thus, microwaving pe rformed in between the first and second staining cycles permits double indi rect immunofluorescence staining with antibodies raised in the same species . Moreover, microwaving also inhibits reactions with endogenous immunoglobu lins present in extracellular compartments. This substantially reduces back ground in indirect immunostaining of mouse tissues with mouse monoclonal an tibodies.