Immunohistochemistry Resource Guide

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Enzyme Substrate Properties

See also Enzyme Substrate Combinations for multiple antigen labeling (p. 30-33) and Counterstain/Substrate Compatibility (p. 35).

Substrate Color Catalog Number Microscopy Mounting Contrast in Pigmented Tissue Multiple Labeling Heat Resistant*
Bright- field Darkfield Electron Fluore- scence Spectral Imaging
Vector® DAB Brown SK-4100 Non-aqueous or Aqueous
Vector® DAB +Ni Gray-Black SK-4100 Non-aqueous
ImmPACT™ DAB Brown SK-4105 Non-aqueous or Aqueous
ImmPACT™ DAB EqV Brown SK-4103 Non-aqueous or Aqueous
Vector® VIP Purple SK-4600 Non-aqueous
ImmPACT™ VIP Purple SK-4605 Non-aqueous
Vector® SG Blue-Gray SK-4700 Non-aqueous or Aqueous
ImmPACT™ SG Blue-Gray SK-4705 Non-aqueous or Aqueous
Vector® NovaRED™ Red SK-4800 Non-aqueous
ImmPACT™ NovaRED™ Red SK-4805 Non-aqueous
Vector® AEC Red SK-4200 Aqueous
ImmPACT™ AEC Red SK-4205 Aqueous
ImmPACT™ AMEC Red Red SK-4285 Aqueous
TMB Blue SK-4400 Non-aqueous
Alkaline Phosphatase
Vector® Red Magenta SK-5100 Non-aqueous or Aqueous
ImmPACT™ Vector® Red Magenta SK-5105 Non-aqueous or Aqueous
Vector® Blue Blue SK-5300 Non-aqueous or Aqueous
Vector® Black Brown-Black SK-5200 Non-aqueous
BCIP/NBT Indigo SK-5400 Non-aqueous or Aqueous

* Substrates that are designated “heat resistant” were developed on tissue then subjected to heat induced epitope retrival (HIER) using a pressure cooker technique (stained tissue was pressure cooked for 1 minute in Antigen Unmasking Solution, returned to room temperature, and rinsed in buffer). Resulting sensitivity after this treatment was found to be equivalent to non-HIER treated tissue.

Alkaline Phosphatase Substrates

Prostate: Prostate Specific Antigen (m), ImmPRESS™ Reagent (HRP), ImmPACT™ DAB (brown).

Tonsil: Cytokeratin AE1/AE3 (m), ImmPRESS™ Reagent (HRP), Vector® DAB-Ni (gray-black).

Tonsil: Cytokeratin AE1/AE3 (m), Vector® ImmPRESS™- AP Reagent, ImmPACT™ Vector® Red (magenta).

Tonsil: Cytokeratin AE1/AE3 (m), ImmPRESS™ Reagent (HRP), ImmPACT™ SG (blue-gray).

Tumor: Cytokeratin (s), VECTASTAIN® Elite® ABC Kit, TMB (blue).

Tonsil: Cytokeratin AE1/AE3 (m), ImmPRESS™-AP Reagent, Vector® Blue (blue).

Tonsil: Cytokeratin AE1/AE3 (m), ImmPRESS™ Reagent (HRP), ImmPACT™ NovaRED™ (red).

Prostate: Prostate Specific Antigen (m), ImmPRESS™ Reagent (HRP), ImmPACT™ AEC (red).

Colon Carcinoma: Pan-Cytokeratin (m), VECTASTAIN® ABC-AP Kit, Vector® Black (brown-black).

Tonsil: CD20 (m), ImmPRESS™ Reagent (HRP), ImmPACT™ VIP (purple).

Tonsil: LCA (m), ImmPRESS™ Reagent (HRP), ImmPACT™ AMEC Red (red).

Prostate: Prostate Specific Antigen (m), VECTASTAIN® ABC-AP Kit, BCIP/NBT (indigo).