
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
Immunohistochemistry Selection Guide
Follow the simple steps below to choose the most appropriate immunohistochemistry (IHC) reagents for your experiment.

Choose Primary Antibody
• Specific for antigen of interest
• Consider tissue species and preparation (fixation)
• Consider antigen retrieval requirements
• Choices determined by the options selected in Steps 1-4
• BLOXALL® Endogenous HRP and AP Blocking Solution
• Avidin/Biotin Blocking Kit (if using VECTASTAIN® ABC system)
• Normal Sera (from the species of secondary antibody)
• M.O.M.® Mouse Ig Blocking Reagent
• Animal-Free Blocker® and Diluent
• BSA
• Casein Solution

Choose Secondary Antibody and Tertiary Detection System
• Choose HRP or AP enzyme system
• Consider sensitivity requirements of immunohistochemistry reagents
• Consider species of primary antibody
• Consider tissue species
OPTION A
OPTION B
OPTION C
One Step
Convenient. Consistent. Ready-to-use.
Non-Biotin based.
• ImmPRESS® Polymer Reagents
OR
Two Step
Economical. Biotin-based.
• Biotinylated secondary antibody + ABC Complex (VECTASTAIN® Elite® ABC Kit)
OR
Two Step
Highest sensitivity. Non-biotin based.
• ImmPRESS® Excel Amplified Polymer Staining Systems

Choose Enzyme Substrate
• Color
• Compatibility with other system reagents (counterstains, mounting media and other substrates for multiplexing)


Choose Mounting Media
• Aqueous vs. non-aqueous
• Compatibility with substrate(s), counterstain, and other immunohistochemistry reagents

Visualize
• Brightfield microscope
Small bowel: CD10 (m), VECTASTAIN® Elite® ABC Kit, ImmPACT® DAB HRP Substrate (brown). Hematoxylin QS counterstain (blue).

Primary Antibody

Secondary Antibody

Amplifier Antibody

Enzyme (HRP or AP)

Biotinylated enzyme (HRP or AP)

Avidin