Immunohistochemistry Resource Guide

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Enzyme Substrate Properties

See also Enzyme Substrate Combinations for multiple antigen labeling and Counterstain/Substrate Compatibility.

Substrate Color Catalog Number Microscopy Mounting Contrast in Pigmented Tissue Multiple Labeling Heat Resistant*
Bright- field Darkfield Electron Fluore- scence Spectral Imaging
Peroxidase
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.

Peroxidase
Substrates
 
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).