Immunohistochemistry Resource Guide

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Counterstain/Substrate Compatibility Table

This table is designed as a reference to determine the optimal counterstain/substrate combination for your application. Considerations should be given to tissue type, antigen unmasking protocol and other detection parameters to achieve the desired staining intensity.

SubstrateCatalog NumberVector® Hematoxylin and Hematoxylin
QS H-3401 and H-3404
Vector® Methyl Green H-3402Vector® Nuclear Fast Red H-3403
ImmPACT™ DAB (brown) SK-4105 Excellent Contrast Excellent Contrast Fair Contrast
ImmPACT™ DAB EqV SK-4103 EXcellent Contrast EXcellent Contrast Fair Contrast
Vector® DAB (brown) SK-4100 EXcellent Contrast EXcellent Contrast Fair Contrast
Vector® DAB-Ni (gray-black) SK-4100 EXcellent Contrast Fair Contrast * Good Contrast
ImmPACT™ AEC (red) SK-4205 EXcellent Contrast Counterstain Incompatibility ** Color Incompatibility
ImmPACT™ AMEC Red (red) SK-4285 EXcellent Contrast Counterstain Incompatibility ** Color Incompatibility
Vector® AEC (red) SK-4200 EXcellent Contrast Counterstain Incompatibility ** Color Incompatibility
TMB (blue) SK-4400 Color Incompatibility Counterstain Incompatibility EXcellent Contrast
ImmPACT™ VIP (purple) SK-4605 Fair Contrast EXcellent Contrast Poor Contrast
Vector® VIP (purple) SK-4600 Fair Contrast EXcellent Contrast Poor Contrast
ImmPACT™ SG (blue-gray) SK-4705 Poor Contrast Good Contrast EXcellent Contrast
Vector® SG (blue-gray) SK-4700 Poor Contrast Good Contrast EXcellent Contrast
ImmPACT™ NovaRED™ (red) SK-4805 EXcellent Contrast EXcellent Contrast *** Color Incompatibility
Vector® NovaRED™ (red) SK-4800 EXcellent Contrast EXcellent Contrast *** Color Incompatibility
ImmPACT™ Vector® Red (magenta) SK-5105 EXcellent Contrast EXcellent Contrast Color Incompatibility
Vector® Red (magenta) SK-5100 EXcellent Contrast EXcellent Contrast Color Incompatibility
Vector® Black (black) SK-5200 EXcellent Contrast EXcellent Contrast * EXcellent Contrast
Vector® Blue (blue) SK-5300 Color Incompatibility Good Contrast EXcellent Contrast
BCIP/NBT (indigo) SK-5400 Color Incompatibility EXcellent Contrast * EXcellent Contrast

* This substrate shows a slight decrease in sensitivity following the methyl green protocol. This decrease can be minimized by reducing the heat incubation and acetone rinse times in the methyl green protocol.
** Substrate dissolves in acetone wash.
*** A slight color change in ImmPACT™ NovaRED™ and Vector® NovaRED™ reaction product may be seen using methyl green.

ProductMountantsCatalog Number
Vector® Hematoxylin Non-aqueous and Aqueous H-3401
Vector® Hematoxylin QS Non-aqueous and Aqueous H-3404
Vector® Methyl Green Non-aqueous H-3402
Vector® Nuclear Fast Red Non-aqueous and Aqueous H-3403
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