StayRed/AP 30ml (Alcohol & Xylene Compatible)

StayRed/AP is a substrate chromogen system designed to be used for either IHC or ISH when using alkaline phosphate detection. StayRed/AP produces a brilliant dark red color. It is also insoluble in organic solvents so sections can be dehydrated in alcohol, cleared in xylene (or a xylene substitute) and permenently mounted.

Intended Use:

Substrate/chromogen in conjunction with alkaline phosphatase-based immunostaining or in situ hybridization systems.

Working Solution:

Aliquot 3mL of StayRed/AP Substrate Buffer in a mixing bottle. Add one drop (~20μL) of StayRed/AP Chromogen. Replace tip, mix, and allow solution to reach room temperature before using.

Note: The working chromogen-substrate solution should be prepared fresh and used within 20-30 minutes of preparation. Any solution not used during this period should be discarded.

Procedure:

• After SA-alkaline phosphatase incubation, wash tissue sections with wash buffer.
• Wipe slides, removing excess buffer. Add enough drops of StayRed/AP working solution to cover tissue sections.
• Incubate for 5-15 minutes at room temperature. For optimal results, observe reaction under microscope for signal development. Once desired signal to noise ratio is achieved, stop reaction by rinsing slides in wash buffer.
• Counterstain sections in hematoxylin.
• Dehydrate sections in alcohol, clear in xylene (or xylene-substitute), and mount in permanent mounting medium.