Fast Red Tablets
Fast Red Tablet Chromogen
Chromogen for Alkaline phosphatase, used for Immunohistohemistry (IHC)
Alkaline Phosphatase based immunostaining systems are gaining wide acceptance among anatomical pathologists because of their relatively high signal to noise ratio. Fast-Red/Napthol AS-Mx, the binary chromogenic substrate for alkaline phosphatase, is the most commonly used substrate chromogen. Napthol AS-Mx acts as the substrate for alkaline phosphatase, and the Fast Red chromogen precipitates at the enzymatic sites producing a vibrant pink color.
Fast Red Chromogen can be used in immunohistochemical and in situ hybridization procedures that employ alkaline phosphatase labeled conjugates.Each Biømeda Fast Red tablet contains 5 mg of Fast Red TR Salt to make 5 mL of working chromogen solution. Fast Red working solution is unstable therefore it is recommended to prepare fresh working solution for each application.Slides should be counterstained with Biømeda Aqueous Hematoxylin (Cat. Np. M10) and mounted in Crystal/Mount (Cat. No. M02). Fast Red is not compatible with organic solvents.
Refrigerate at 4°C. Do not freeze.
Working Chromogen Preparation: Remove the Fast Red Chromogen tablets from the refrigerator 20 minutes before the preparation of the working chromogen solution. This will minimize the condensation of ambient water on the tablets during handling.
1.Add one tablet to 5 mL of the freshly prepared chromogen substrate solution. The composition of chromogen substrate is: 50 mM Tris, pH 8.0, 75 mM sodium chloride, 5 mM magnesium chloride, 1 mM ZnCl2, 0.01% of Naphtol AS-Mx Phosphate (predissolved Naphtol in 100 microliters of 50% Dimethylformamide) in distilled or deionized water.
2.Allow tablet to dissolve for approximately 1 minute with occasional mixing. Secure the cap of the bottle containing the unused tablets and return it to the original plastic box.
3.Filter the working chromogen solution using syringe filter (optional).
4.Apply this solution to the specimens.
5.Incubate for 5-10 minutes at room temperature. Wash specimens with distilled water and proceed with the counterstaining steps.
Fast red chromogen substrate can be mounted with aqueous mounting medium. We in our lab use Crystal mount (M02) for mounting slides.
THIS CHROMOGEN CAN NOT WITHSTAND DEHYDRATION WITH ORGANIC SOLVENTS AND SHOULD BE MOUNTED WITH AQUEOUS MOUNTING MEDIA.
Use the working chromogen solution within 2 hours from the time of the addition of the tablet. Discard unused reagent according to local and state regulations.