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Regarding the MULTIVIEW® PLUS (mouse-HRP/mouse-AP) IHC kit (Brown/Green), sufficient washes between antibody, detection, and chromogen steps (3 x 1 minute in wash buffer) will remove excess antibody, detection reagent, or chromogen. Antibody removal is therefore not necessary. Also, chromogens of the first layer may act as blocking chromogens, thus shielding the first layer from subsequent reactions. DAB is a good shielding chromogen, and the antibody removal can often be skipped when the chromogen of the first layer is DAB.