Immunohistochemistry Resource Guide

Choose Primary Antibody

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

VECTABOND® Reagent
(Tissue Section Adhesive)

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 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 Nuclear Counterstain

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

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).