Immunohistochemistry Resource Guide

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IHC Guide Cover v1

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

Mounting Media

VectaMount® Permanent Mounting Medium is an optically clear formula for permanently preserving histochemical stains or precipitable enzyme substrates in tissue sections or cell preparations.

  • Non-aqueous
  • Toluene- and xylene-free (low toxicity)
  • Resin-based media
  • Odorless
  • Viscosity formulated for easy application and uniform spreading
  • Compatible with most horseradish peroxidase and alkaline phosphatase substrates
  • Dries clear with an ideal refractive index suitable for high-resolution oil-immersion microscopy

VectaMount AQ Aqueous Mounting Medium

VectaMount AQ Aqueous Mounting Medium preserves the color and clarity of enzyme substrates whose reaction products are soluble in alcohol or other organic solvents. Stained and mounted sections can be stored in a slide box at room temperature for at least two years without fading.

  • Hard-setting
  • Simple to use, requires no mixing

Product

Catalog Number

VectaMount® Permanent Mounting Medium

H-5000-60

VectaMount® AQ Aqueous Mounting Medium

H-5501-60

Mounting Media / Substrate Compatibility

Substrate

VectaMount® Permanent
Mounting Medium

VectaMount® AQ Aqueous
Mounting Medium

HRP

Vector® DAB

•

•

Vector® DAB-Ni

•

ImmPACT® DAB

•

•

ImmPACT® DAB EqV

•

•

Vector® VIP

•

ImmPACT® VIP

•

Vector® NovaRED®

•

ImmPACT® NovaRED®

•

Vector® SG

•

•

ImmPACT® SG

•

•

Vector® AEC

•

ImmPACT® AEC

•

ImmPACT® AMEC Red

•

Vector® TMB

•

AP

Vector® Red

•

•

ImmPACT® Vector® Red

•

•

Vector® Blue

•

•

Vector® Black

•

Vector® BCIP/NBT

•

•

image 98
Melanoma: S100 (rp), VECTASTAIN® Elite® ABC Kit, Vector VIP (purple), Vector Methyl Green counterstain (green). Note color contrast with brown pigments in tissue.
image 99
Cytospin of EBV+ cell line: Epstein-Barr virus nuclear antigen 1 (EBNA-1; rat), ImmPRESS Reagent (HRP) Anti-Rat IgG, ImmPACT NovaRED (red). (Image courtesy of Dr. GM Reynolds, Centre for Liver Research, University of Birmingham, U.K.)
image 100
Breast carcinoma: • CD31 (m), ImmPRESS Anti-Mouse IgG HRP Reagent, ImmPACT DAB EqV HRP Substrate (brown) • MRC1 (r), ImmPRESS- AP Anti- Rabbit IgG Reagent, ImmPACT Vector Red AP Substrate (magenta). Image courtesy of Richard Allen, part of the Academic Unit of Inflammation and Tumour Targeting headed by Professor Claire Lewis.
image 101
Colon: • CD3 (rm), ImmPRESS® Reagent (HRP) Anti-Rabbit IgG, ImmPACT SG (blue-gray) • Cytokeratin AE1/AE3 (m), ImmPRESS Reagent (HRP) Anti-Mouse IgG, ImmPACT AMEC Red (red).
image 102
Tumor: • CD34 (m), VECTASTAIN Universal ABC-AP Kit, Vector Blue AP Substrate (blue) • Cytokeratin 8/18 (m), VECTASTAIN Universal ABC-AP Kit, Vector Red AP Substrate (red).
image 103
Melanoma: Vimentin (rm), ImmPRESS-AP Anti-Rabbit IgG Reagent, Vector Blue AP Substrate (blue). Note color contrast with brown pigments in tissue.

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