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Immunohistochemistry Resource Guide

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Table of Contents

Immunohistochemistry Selection Guide

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

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Choose Primary Antibody

• Specific for antigen of interest
• Consider tissue species and preparation (fixation)
• Consider antigen retrieval requirements

SECONDARY DETECTION SYSTEMS

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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

2 optionA 2-optionA 2-optionA
OPTION A
2 optionB 2-optionB 2-optionB
OPTION B
2 optionC 2-optionC 2-optionC
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

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Choose Enzyme Substrate

• Color

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

3 Enzyme Substrate 3-Enzyme_Substrate 3-Enzyme_Substrate
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Choose Nuclear Counterstain

• Blue, green, or red

• Compatibility with substrate,
mounting media

4 Nuclear Counterstain 4-Nuclear_Counterstain 4-Nuclear_Counterstain
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Choose Mounting Media

• Aqueous vs. non-aqueous

• Compatibility with substrate(s), counterstain, and other immunohistochemistry reagents

5 Mounting Media 5-Mounting_Media 5-Mounting_Media
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Visualize

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

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