Biotinylated HRP and biotinylated AP are most commonly used in immunohistochemistry (IHC) to amplify the signal of biotinylated primary antibodies using the ABC staining method. Biotinylated Fluorescein (FITC) cannot be used to polymerize and amplify the signal to the same degree as biotinylated enzymes because each fluor molecule is tagged with only one biotin molecule. However, the fluorescent variant of traditional ABC staining is possible with this unique molecule.
Biotinylated enzymes – biotin-labeled horseradish peroxidase (B-HRP), alkaline phosphatase (B-AP) and beta-galactosidase (B-bGal) for use in avidin-biotin complex (ABC) staining
Biotinylated FITC – amplify the fluorescein signal of biotinylated primary antibodies by using avidin or streptavidin plus biotinylated fluorescein (B-FITC)
Biotinylated controls – use biotinylated bovine serum albumin (B-BSA) or any one of these molecules as positive controls in biotin quantitation methods