AEC Chromogen/Substrate Kit Protocol (NB900-79773): https://www.novusbio.com/products/aec-chromogen-substrate_nb900-79773
1. Add 5 ml of distilled or de-ionized water to a test tube.
2. Add two drops of concentrated buffer and mix.
3. Add two drops of concentrated AEC chromogen and mix.
4. Add two drops of 3% H2 O2 substrate solution and mix.
Working chromogen solution is stable for 6 hr. Any solution not used after this period should be discarded.
1. Once tissue sections have been incubated with peroxidase, wash them with buffer thoroughly.
2. Wipe the glass to remove excess of buffer and add enough of the AEC working solution to cover the tissue sections.
3. Incubate for 10-20 minutes at room temperature. For best results, observe signal development under the microscope. Once desired signal to noise ratio is achieved, stop the reaction by washing the slides in wash buffer.