Amylase Solution

Amylase Solution

Assay-kits

Product code: ACMBS-38
Specifications: 20mL, 100mL (Box)
Intended use: Intended for identifying glycogen and other polysaccharides
Store temperature: 2℃～8℃
Expiry date: 12 Months

More Details

PrincipleMain componentsSample requirementTest procedureResult interpretationMethod limitationPrecautions

Principle

Amylase is a generic term for enzymes that hydrolyze starch and glycogen. Amylase generally acts on soluble starch, amylose, glycogen, etc., and cuts off the α-1,4-chain indiscriminately, and decomposes it into disaccharides and monosaccharides. Since monosaccharides are extracted during histochemical operations such as immobilization, dehydration, and embedding, the sugars that can be displayed on general tissue specimens are mainly polysaccharides, including glycogen, mucopolysaccharides, mucins, glycoproteins, glycolipid and so on. Therefore, to determine whether the red substance after PAS staining is glycogen, it is also necessary to carry out amylase digestion comparison experiments.

Main components

Reagent composition	Main ingredient
Amylase solution	α-Amylase

Sample requirement

Specimens should be fresh tissue section or cell smears, and the tissue section should be adequately fixed.

Test procedure

Cell smear staining:

1. Two consecutive smears were taken from the same patient and identified as A and B respectively. After natural drying, they were fixed with formaldehyde vapor or fixed solution (purchased separately) for 1 minute, and washed by distilled water.
2. The tissue of slice B was covered with amylase solution and digested in 37℃ incubator.
3. For 40 minutes.
4. Wash the slice B by running water, and then by distilled water.
5. Add periodic acid solution (purchased separately) on A and B tablets simultaneously for 10～15 minutes, and washed by distilled water.
6. Drop Schiff reagent (purchased separately) 10 minutes, and washed by distilled water.
7. Add Hematoxylin stain (purchased separately) or Methyl green Solution (purchased separately) to stain it for 2～5 minutes, then washed by running water. Microscopic examination after drying.

Tissue section staining:

1. Two consecutive smears were taken from the same patient and identified as A and B respectively. Dewaxing to water, and washed by distilled water.
2. The tissue of slice B was covered with amylase solution and digested in 37℃ incubator for 40 minutes.
3. Wash the slice B by running water, and then by distilled water.
4. Add periodic acid solution (purchased separately) on A and B slices simultaneously (purchased separately) for 10 minutes, and washed by distilled water.
5. Drop Schiff reagent (purchased separately) 10～15 minutes, and washed by distilled water.
6. Add Hematoxylin stain (purchased separately) to stain it for 2～3 minutes, then washed by running water, returned blue for 3-5 minutes.
7. Dehydrate in typical manner and clear. Mount with mounting media.

Result interpretation

A red to reddish purple reaction in the cytoplasm of slice A without amylase digestion was positive for PAS staining. If slice B was negative after digestion, the positive substance of slice A was glycogen, if slice B was still positive after digestion and staining, it showed that the positive substance of the slice A was other polysaccharides.

Method limitation

Only used for observation of tissue and cellular content staining.

Precautions

1. The kit recovery at room temperature before use and shake the reagent before using.
2. When incubating at 37℃ in a warm box, attention should be paid to avoiding volatilization and drying of amylase solution.
3. Interpretation of staining results should be made by well-trained technicians and pathologists.
4. Should carefully read the instruction manual before using. Can only be used before expiration date, and good personal hygiene protection.
5. Dispose of waste according to the requirements of the hospital or environmental protection department after use.
6. The production date, production batch number and expiration date on the package.

References

1. Chinese Medical Association. Clinical technical operating specifications-pathology volumes [M]. People's Military Medical Publishing House.

2. Ling Qibo. Practical pathology special staining and histochemistry technology [M]. Guangdong Higher Education Press.

3. People's Republic of China national health and Family Planning Commission of medical affairs authority. The clinical laboratory operating procedures [M]. Fourth edition. Beijing: People's Medical Publishing House, 2014.

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