Immunohistochemistry Resource Guide

Table of Contents

Introduction

Immunohistochemistry Workflow

Immunohistochemistry Selection Guide

Pioneering in IHC Technology

Choosing a Detection System

Avidin-Biotin Complex (ABC)-Based Detection

VECTASTAIN^® ABC Systems

VECTASTAIN^® ABC Kits

Choosing a VECTASTAIN^® ABC Kit

> Customizing your VECTASTAIN^® ABC Kit

Polymer-Based Detection

ImmPRESS^® One-Step Polymer Systems

> ImmPRESS^® Two-Step Amplified Polymer Systems

> Choosing an ImmPRESS^® Polymer Kit

> Multiple Antigen Labeling Simplified

Species on Species Detection

M.O.M.^® (Mouse on Mouse) Immunodetection Kits

Choosing an Enzyme Substrate

Enzyme Substrates

> Enzyme Substrate Properties

Multiple Antigen Labeling

Enzyme Substrate Combinations

Counterstaining

> Counterstain/Substrate Compatibility Table

Blocking Background Signal

Secondary and Tertiary Detection Reagents

Mounting Media

Accessory Reagents

Counterstaining

A counterstain introduces color to specific cellular structures to provide contrast to the colored enzyme substrate. Counterstaining aids in visualization and target localization, facilitating interpretation of morphology and cell structure within the tissue section. Our nuclear counterstains are packaged as convenient, ready-to-use solutions for use on individual slides or in staining dishes. Vector^® Hematoxylin (blue)

• Based on Gill’s III formulation
• Progressive stain formula. The intensity can be adjusted to optimize results for either manual or automated systems
• Excellent color contrast with most commonly used peroxidase and alkaline phosphatase substrates
• Suitable for use with non-aqueous and aqueous mounting media
• Alcohol- and mercury-free

Tumor tissue section showing specific cytoplasmic cell staining (Vector DAB, brown) with Vector Hematoxylin QS counterstain (blue).

Vector Hematoxylin QS (blue)

• Modification of Mayer’s hematoxylin developed especially forimmunocytochemistry
• Ready-to-use without filtration or ‘blueing’ step
• Stains in less than 45 seconds
• Excellent color contrast with most commonly used peroxidase and alkaline phosphatase substrates
• Suitable for use with non-aqueous and aqueous mounting media
• Mercury-free

Methyl Green (light green)

• Superior formulation of methyl green suitable for use with a wide range of enzyme substrates
• Simple, two step procedure
• Excellent alternative in multiple antigen labeling when hematoxylin obscures the substrate colors
• Suitable for use with non-aqueous mounting media

Vector Nuclear Fast Red (pink)

• Fast one-step protocol
• Excellent alternative in multiple antigen labeling when hematoxylin obscures the substrate colors
• Good contrast with a variety of substrates

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