Technovit H7100 / H8100 Acid Phosphatase
EMS Catalog # 14653-14654
Acid Phosphatase For Glycol Methacrylate Sections
Procedure
- Incubate sections in the incubating medium at 37°C for five to 12 hours. Long incubation periods are needed to get significantly visible reaction product.
- Wash in distilled water for two minutes.
- Counterstain with Methyl Green for five minutes.
- Wash in distilled water for two minutes.
- Air dry and cover slip.
Results
Nuclei |
dark green |
Cytoplasm |
light green |
Sites of enzyme activity |
red |
Solutions
Incubating Medium:
- Combine 20 ml of buffer solution, 48 ml of distilled water and 4 ml of substrate solution.
- Combine 3.2 ml of Pararosaniline solution with 3.2 ml of sodium nitrite solution. Mix for one minute.
- Add the second solution to the first.
- Adjust pH to 5.
Buffer Solution:
- 5.9 g Anhydrous sodium acetate
- 14.7 g Sodium barbiturate
- 500 ml Distilled water (boiled)
Do not adjust the pH of the buffer and store at 4°C.
Substrate solution:
- 40 mg Naphtol As-BI phosphatease, sodium salt
- 4 ml N.N-dimethylformamide
Pararosaniline Solution:
- 2 g Pararosaniline (C.I.#42500)
- 50 ml 2N HCl
Use heat to dissolve, filter when cool and store at 4°C
Sodium Nitrite Solution:
- Sodium Nitrite - 1 gm
- Distilled Water - 25 ml
- Prepare fresh and store at 4°C.
Methyl Green
- Methyl Green (C.I.# 42585) - 1 g
- Phosphate/citrate buffer 0.1M pH 4.0 - 100 ml
Citation:
- Gerrits, P. O. and Smid, L., "Staining Procedures for Tissues Embedded in 2-Hydroxyethyl Methacrylate", Heraeus Kulzer.
Product Information
Technovit Glycol Methacrylate embedding kits H7100 / H8100