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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:

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

Buffer Solution:

  • 2.4gm 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 0.2gm of Nuclear Fast Red add 200ml of boiling 0.5% aluminum sulfate solution
  • Keep boiling for five to ten minutes.
  • Allow to cool and filter before use.

Online Ordering

Technovit Glycol Methacylate Kits H7100 / H8100