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.

Substrate Catalog Number Vector® Hematoxylin and Hematoxylin
QS H-3401 and H-3404
Vector® Methyl Green H-3402 Vector® Nuclear Fast Red H-3403
ImmPACT® DAB EqV SK-4103 Excellent Contrast Excellent Contrast Fair Contrast
ImmPACT® DAB (brown) SK-4105 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.

Product Mountants Catalog 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