Mixing of Reagent:
- To 4.9 ml of distilled or deionized water in a test tube, add five drops of buffer reagent B, mix well.
- Add Fast Red tablet (Reagent C), let stand in this solution for 5-10 minutes. Vortex for 3-5 minutes till the entire tablet is dissolved.
- Add 5 drops of substrate reagent S, mix.
(This ready to use reagent is good for several hours. The unused solution can be discarded according to city, county, state, province or country’s regulations).
Application: IHC, ICC
- After alkaline phosphatase reagent, wash slide with buffer and distilled/deionized water.
- Add few drops of ready to use Fast Red reagent; incubate at RT or 37ºC for 5-10 minutes.
The optimum time of incubation for Fast red Super should be determined by the individual lab.
- Wash slides 2-5X with buffer, followed by rinsing with distilled/deionized water.
- Add counterstain compatible with red color. We use hematoxylin for 30-60 seconds, followed by washing with tap water, buffer with pH over 7 to convert purple blue color to blue color.
- The slide can be mounted with aqueous mounting medium. See #6 or organic mounting medium see #7).
- Wash the slide with distilled/deionized water. The slide can be mounted with aqueous mounting medium (we use ImmunoHistoMount cat. # AR-6503; see data sheet).
- Or the slide can be mounted with organic mounting medium. (Dehydrate with increasing % of alcohol, 100% alcohol, followed by limonene, toluene, xylene or any other dehydrating reagent.. (We use Organo/limonene mounting medium cat. # AR-6504, please follow data sheet for mounting medium).