The HIGHDEF® Red IHC Chromogen can be used in conjunction with peroxidase-based immunostaining systems. Peroxidase from the immunodetection system reacts with hydrogen peroxide substrate to degrade it, which reacts with AEC, precipitating it at positive sites yielding a reddish brown colored product.
HIGHDEF® Red IHC Chromogen is a single, highly stable, AEC chromogen/substrate working solution. When used in conjunction with immunoperoxidase detection systems, AEC produces a red colored end product at positive sites, yielding strong contrast when combined with a blue hematoxylin counterstain. AEC has been well accepted among pathologists because it is less chemically hazardous than DAB. Specimens stained using HIGHDEF® Red IHC Chromogen cannot be dehydrated in ethanol and must be mounted in an aqueous-based mounting medium such as HIGHDEF® IHC mount (Prod. no. ADI-950-261).
Protocol:
Once sections have been incubated with peroxidase, wash with wash buffer.
Blot slides to remove excess buffer. Add enough drops of HIGHDEF® Red IHC Chromogen to cover tissue sections.
Incubate for 5-15 minutes at room temperature. For best results, observe reaction under a microscope for signal development. Once desired signal-to-noise ratio is achieved, stop the reaction by washing slides in DI H2O.
Regulatory Status:
RUO - Research Use Only
Immunohistochemistry analysis: Formalin fixed paraffin embedded human tonsil stained with a high molecular weight CK antibody labeled with HIGHDEF® IHC substrate (AEC, stable) (Prod. no. ADI-950-210) produces a robust red color.
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Product Literature References
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