Fast Red Tablets (CAS 3165-93-3)

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Fast Red Tablets is a substrate for alkaline phosphate (AP) based detection systems. Each tablet contains 0.5 mg naphthol substrate, 2 mg Fast Red chromogen and 0.4 mg levamisole, an inhibitor of endogenous AP activity. The compound is suitable for fluorescent microscopy, immunoblotting, immunohistochemistry, in-situ hybridization and flow cytometric assays. Fast Red Tablets are recommended for use with Crystal Mounting Medium (sc-24943).
In an improved in-situ hybridization assay the Fast Red Tablets were incorporated to detect a DIG labeled probe through fluorescent detection of AP activity. With the use of bright field microscopy and laser scanning microscopy Fast Red Tablets have been used to detect HPV DNA from cell deposits or tissue sections. To study the effects of glucocorticoid dexamethasone on cell proliferation and AP expression in rat bone marrow stromal cells Fast Red Tablets were used to detect enzymatic activity in whole cells quantified by flow cytometry.

To prepare the substrate solution:
• Dissolve 1 Fast Red tablet in 2 ml 0.1 M Tris-HCl, pH 8.2.
• Shake for 1-3 minutes. Some particles will not dissolve, but that will not affect the experimental results.
• Use substrate solution within 30 minutes.

To use in immunohistology:
• Cover the tissue section with between 0.1 and 0.2 ml of the Fast Red solution.
• Fast Red is a fast reacting substrate and experiments should be monitored during the reaction to prevent overdevelopment. Reaction may be stopped by gently washing the slide in water.
• If haziness of the Fast Red solution occurs, it can be removed by running the solution through a 0.2 µm filter.

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