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EMS Technical Data Sheets
Technovit H7100 / H8100 Alkaline Phosphatase
EMS Catalog #14653-14654
Alkaline Phosphatase For Glycol Methacrylate Sections
Procedure
- Incubate sections in the incubating medium at room temperature for one to three hours. Two hours is sufficient in most cases.
- Wash in distilled water for two minutes.
- Counterstain with Nuclear Fast Red for five to ten minutes.
- Wash in distilled water for two minutes.
- 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.