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