Immunohistochemistry Resource Guide

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Immunohistochemistry Selection Guide

Follow the simple steps below to choose the most appropriate labeling and detection solution for your experiment.

Choose Primary Antibody

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

Blocking Reagents
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 Block and Diluent
BSA
Casein Solution

SECONDARY DETECTION SYSTEMS

Choose Secondary Antibody and Tertiary Detection System

• Choose HRP or AP enzyme system
• Consider sensitivity requirements
• 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 Nuclear Counterstain

• Blue, green, or red
• Compatibility with substrate,
mounting media

Choose Mounting Media

• Aqueous vs. non-aqueous
• Compatibility with substrate(s)
and counterstain

Visualize

• Brightfield microscope


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

Legend

Primary Antibody

Secondary Antibody

Amplifier Antibody

Enzyme (HRP or AP)

Biotinylated enzyme (HRP or AP)

Avidin
3