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Electron Microscopy Sciences

Technical Data Sheets

Technovit H7100 / H8100 Alkaline Phosphatase

EMS Catalog #14653-14654

Alkaline Phosphatase For Glycol Methacrylate Sections

Procedure

  1. Incubate sections in the incubating medium at room temperature for one to three hours. Two hours is sufficient in most cases.
  2. Wash in distilled water for two minutes.
  3. Counterstain with Nuclear Fast Red for five to ten minutes.
  4. Wash in distilled water for two minutes.
  5. Air dry and coverslip.

Results

Nuclei red
Sites of enzyme activity blue

To preserve the reaction product, selection of the right mounting (coverslip) medium is important.

Solutions

Incubating Medium:

  • 5 mg Naphtol As-MX phosphate, di-sodium salt (sigma)
  • 0.25 ml N,N-dimethylformamide
  • 30 mg Fast blue BB (sigma)
  • 25 ml Distilled water
  • 25 ml Buffer Solution
  • 2 drops 10% Magnesium Sulfate Solution
    Prepare fresh, shake well and filter before use.

Buffer Solution:

  • 2.4 g 0.2M Tris (Hydroxyethyl)-aminomethane
  • 100ml Distilled water
  • Adjust the pH of the buffer to 8.9 with dilute HCl and store at 4°C.

Nuclear Fast Red:

  • To 0.2 g of Nuclear Fast Red add 200 ml of boiling 0.5% aluminum sulfate solution
  • Keep boiling for five to ten minutes.
  • Allow to cool and filter before use.

Product Information

Technovit Glycol Methacylate Kits H7100 / H8100