Peroxidase chromogen-substrate 3-amino-9-ethyl-carbazole (AEC) is an excellent chromogen system that is used in Immunohistochemistry (IHC) and Immunocytochemistry (ICC).
- To 5 ml of distilled or deionized water in a test tube, add two drops of reagent B buffer, mix well.
- Add one drop of reagent C chromogen, mix well.
- Add one drop of reagent S substrate, mix well. This ready to use reagent is good for several hours. The unused AEC solution can be discarded according to city, county, state, province or country’s regulations.
- After the peroxidase reaction, wash slide with buffer (this buffer should not contain any sodium azide because peroxidase is inactivated), and distilled/deionized water.
- Add few drops of ready to use AEC reagent, incubate at RT or 37ºC for 5-10 minutes.
- Wash 2-5X with buffer, followed by rinsing with distilled/deionized water.
- Add counterstain compatible with AEC, we use hematoxylin 30-60 seconds (cat. # AR-6521).
- Wash with tap water, buffer pH 7.4 or higher followed by rinsing with distilled/deionized water.
- Mount slide with aqueous mounting medium, we use our IHM (cat. # AR-6503) please follow the protocol for mounting medium.