BioGnost’s P.A.S. Diastase kit is most commonly used for identifying glycogen in liver. Periodic acid enables the molecules containing glycol groups to create aldehydes affected by Schiff’s reagent staining them violet (magenta). Specific stains are created by applying the PAS method on unsubsti-tuted polysaccharides, mucoproteins and glycoproteins, glycolipids and phospholipids. Alpha-amylase enzyme (also known as diastasis) is used for differentiation between glycogen and other PAS-positive structures by dissolving 1→4 glycosidic bonds, causing the glycogen to remain unstained after the PAS reaction. BioGnost’s P.A.S. Diastase kit uses thermostable enzyme which does not require heating to +37°C to be active, but incubat-ing the section at +37°C is preferred in order to achieve better glycogen breakdown. The same tissue section is used as negative control for this reaction, but the sample is not treated using alpha-amylase.
P.A.S. Diastase Kit
BioGnost’s P.A.S. Diastase kit is most commonly used for identifying glycogen in liver. Periodic acid enables the molecules containing glycol groups to create aldehydes affected by Schiff’s reagent staining them violet (magenta). Specific stains are created by applying the PAS method on unsubsti-tuted polysaccharides, mucoproteins and glycoproteins, glycolipids and phospholipids. Alpha-amylase enzyme (also known as diastasis) is used for differentiation between glycogen and other PAS-positive structures by dissolving 1→4 glycosidic bonds, causing the glycogen to remain unstained after the PAS reaction. BioGnost’s P.A.S. Diastase kit uses thermostable enzyme which does not require heating to +37°C to be active, but incubat-ing the section at +37°C is preferred in order to achieve better glycogen breakdown. The same tissue section is used as negative control for this reaction, but the sample is not treated using alpha-amylase.
For 100 tests.
Description
PROCEDURE
Using kit for 100 tests | ||
1 | Deparaffinize the section in xylene (BioClear) or in a xylene substitute (BioClear New) | 3 exchanges, 10 minutes each |
2 | Rehydrate using 100% alcohol (Histanol 100) | 2 exchanges, 5 and 3 minutes |
3 | Rehydrate using 95% alcohol (Histanol 95) | 2 minutes |
4 | Rehydrate in distilled (demi) water | 2 minutes |
5 | Add Hematoxylin G1 to the sections (≥5 drops) | 5 minutes |
6 | Rinse under tap water | 4 minutes |
7 | Immerse the sections in freshly prepared Congo Red working solution | 18 minutes |
8 | Rinse under tap water | Until excessive dye is rinsed (a few minutes) |
9 | Dehydrate using 70% alcohol (Histanol 70) | 10 dips |
10 | Dehydrate using 95% alcohol (Histanol 95) | 10 dips |
11 | Dehydrate using 100% alcohol (Histanol 100) | 3 minutes |
12 | Clear the section in xylene (BioClear) or in a xylene substitute (BioClear New) | 2 exchanges, 5 minutes each |
Immediately after clearing apply an appropriate BioMount medium for covering/mounting on the section. If BioClear xylene was used, use one of BioGnost's mounting xylene-based media (BioMount, BioMount High, BioMount M, BioMount DPX, BioMount C, or universal BioMount New). If BioClear New xylene substitute was used, the appropriate covering agent is BioMount New. Cover the section with VitroGnost cover glass. | ||
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