Thermo Scientific™

Lab Vision™ Liquid Fast-Red Substrate System

 Related applications:
Anatomical Pathology
Employ a widely used chromogen for immunohistochemical staining and immunoblotting with the Thermo Scientific™ Lab Vision™ Liquid Fast-Red Substrate System.

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Description

In the presence of alkaline phosphatase enzyme, Fast-Red produces a bright red reaction product.
  • Bright red reaction product can be seen using brightfield or fluorescent microscopy
  • Liquid Fast-Red is not soluble in xylenes and alcohols; therefore it may be permanently mounted

Staining Protocol:

1. Prepare the working solution immediately before use.

2. Add 1 drop (40μL) of Liquid Fast-Red Chromogen to 3mL of Naphthol Phosphate Substrate and mix it well.

3. Apply solution to tissue section. Use working solution within 15 minutes of preparation or decreased sensitivity may result.

4. Incubate tissue section for 10-25 minutes.

5. Rinse tissue, counterstain, and coverslip.

For permanent cover slip mounting:

a. Air dry slides on 37°C warming plate before applying permanent mounting media (preferred method).

OR

b. Dehydrate and clear quickly through alcohol and xylene (no more than 10 seconds each) before mounting with permanent mounting media.

For aqueous cover slip mounting:

a. Any gelatin-based mounting media (i.e., Permafluor TA-030-FM)

b. UltraMount aqueous mounting media (TA-060-UM), dries hard, for use without a cover slip)

c. VisionMount aqueous mounting media (TA-060-UG), dries hard, for use with a cover slip)

Note: remove UltraMount and VisionMount with alcohol.

Staining Protocol:

1. Prepare the working solution immediately before use.

2. Add 1 drop (40μL) of Liquid Fast-Red Chromogen to 3mL of Naphthol Phosphate Substrate and mix it well.

3. Apply solution to tissue section. Use working solution within 15 minutes of preparation or decreased sensitivity may result.

4. Incubate tissue section for 10-25 minutes.

5. Rinse tissue, counterstain, and coverslip.

For permanent cover slip mounting:

a. Air dry slides on 37°C warming plate before applying permanent mounting media (preferred method).

OR

b. Dehydrate and clear quickly through alcohol and xylene (no more than 10 seconds each) before mounting with permanent mounting media.

For aqueous cover slip mounting:

a. Any gelatin-based mounting media (i.e., Permafluor TA-030-FM)

b. UltraMount aqueous mounting media (TA-060-UM), dries hard, for use without a cover slip)

c. VisionMount aqueous mounting media (TA-060-UG), dries hard, for use with a cover slip)

Note: remove UltraMount and VisionMount with alcohol.



Documents

Manuals & protocols