Immunohistochemistry Resource Guide

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Choosing a Detection System

Immunohistochemistry Overview

Immunohistochemistry (IHC) is a method to detect specific target antigens (proteins) in tissue sections using antibodies. Immunocytochemistry (ICC) uses similar techniques to localize cellular proteins in cell preparations. Both IHC and ICC are powerful tools that provide insights into gene expression, spatial relationships, and biomarker identification in a wide variety of applications. These applications include basic research, assessment of normal and disease states within human and animal tissues, and assessment of plant pathology.

The target antigen, bound by the detection antibody, is visualized using either chromogenic or fluorescence detection. In chromogenic detection, the detection antibody is conjugated to an enzyme. The enzyme, usually horseradish peroxidase or alkaline phosphatase, catalyzes the conversion of its respective chromogen to a colored precipitate at the site of the antigen. This precipitate can be visualized by using brightfield microscopy. Certain chromogens can also be visualized by using electron, darkfield or fluorescence microscopy. In fluorescence detection, the detection antibody is conjugated to a fluorophore which can be visualized using fluorescence microscopy.

For the purposes of this antibody selection guide, IHC will be referenced for both IHC and ICC techniques.

Comparison of Detection Systems and Antibody Selection

Choose the appropriate detection system for your experiment based on enzyme, sensitivity, cost, biotin vs. non-biotin formats, flexibility, and time considerations. These parameters will help you with your antibody selection.

Detection System Enzyme Sensitivity Cost/ Assay Biotin- Free Micro- polymer Modular Mouse Primary on Mouse Tissue Ready-to- Use (R.T.U.) Format Typical number of steps
ImmPRESS™ Kits
ImmPRESS™ Excel Amplified HRP Polymer Kits HRP •••••• ••••     2
ImmPRESS™ HRP Polymer Kits HRP ••••• •••     1
ImmPRESS™ VR HRP Polymer Kits HRP ••••• ••••     1
ImmPRESS™ AP Polymer Kits AP ••••• ••••     1
VECTASTAIN® Elite® ABC Kits HRP ••••• ••         2
R.T.U. VECTASTAIN® Elite® Kits HRP ••••• ••       2
VECTASTAIN® Universal Quick Kits HRP •••• ••         2
R.T.U. VECTASTAIN® Universal Quick Kits HRP •••• ••       2
VECTASTAIN® ABC-AP Kits AP ••••         2
Original VECTASTAIN® ABC Kits HRP •••         2
Mouse on Mouse (M.O.M.™) Kits
Mouse on Mouse (M.O.M.™) ImmPRESS™ Polymer Kit HRP ••• •••     1
Mouse on Mouse (M.O.M.™) Kits HRP ••• •••       2
Additional Options
Enzyme Conjugated Avidin/Streptavidin HRP or AP •••         2
R.T.U. HRP Avidin/Streptavidin HRP •••       2
Enzyme Conjugated Secondary Antibody HRP or AP ••         1

HRP - Horseradish peroxidase
AP - Alkaline phosphatase
VR - Veterinary Reagents