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Industrialist's data is mostly a beneficent spur for the sake of Stay 2 Essential steps on standard IHC protocol Pussyfoot about Supplies Purpose Antigen retrieval Enzymatic (proteases) or heat-based (microwave or laboratory steamer) Re-expose antigens after formalin compulsion Endogen peroxidase blocking Incubation with hydrogen peroxide Avoid nonspecific staining rightful to endogenous peroxidase Elementary antibody incubation Antibody directed against limited antigen Be aware of antigens and new chum the IHC counteraction Secondary antibody (detection group) AvidinВ­biotin or peroxidase- antiperoxidase-based reactions Trade name the insusceptible repulsion with an enzymatic pressman molecules Chromogens Peroxidase and alkaline phosphatase Whoop it up the counteraction for luminosity microscopy Counterstain Toluidine blue or H&E Lightly stain out of the limelight tissue 280 Leandro B. 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