AB-PAS staining kit - BioReagent, for microscopy, Biological Stain, high purity

Cat. No.: A774824

GRADE & PURITY BioReagent, Biological Stain, for Microscopy

Overview

  Alcian blue (also known as Alcian blue, Alcian blue, etc.) and PAS technique can be used together to identify neutral mucin and acidic mucin in the same tissue section. This technique is also commonly used as a means to extensively detect mucin. The section is first stained with standard Alcian blue (pH=2.5) and then subjected to the PAS technique. Alcian blue stains sialomucin, sulfomucin, and proteoglycans blue. The PAS technique stains neutral mucin deep red or reddish-purple, while staining tissues and cells containing both neutral and acidic mucin in varying shades of purple. This is due to the combination and reaction of Alcian blue with Schiff reagent. The above staining often appears in small intestinal goblet cells containing neutral mucin and sialomucin. The staining principle of Alcian blue lies in its being a copper phthalocyanine dye. This cationic dye binds to acidic groups, meaning Alcian blue forms insoluble complexes with anionic groups in tissues, such as carboxyl and sulfate groups. The positively charged salt bonds in the molecule combine with the negatively charged acidic groups in acidic mucopolysaccharides to form insoluble complexes that appear blue. When combined with PAS for composite staining, it can display three different mucus substance components.

A774824	Component	6×50 mL	6×100 mL	Storage
A774824A	3% Acetic Acid Solution	50 mL	100 mL	RT.
A774824B	Alcian Blue Staining Solution	50 mL	100 mL	2-8℃. Store in the dark
A774824C	Periodic Acid Solution	50 mL	100 mL	2-8℃. Store in the dark
A774824D	Schiff Reagent	50 mL	100 mL	2-8℃. Store in the dark
A774824E	Hematoxylin Staining Solution	50 mL	100 mL	2-8℃. Store in the dark
A774824F	Acidic Ethanol Differentiation Solution	50 mL	100 mL	RT.

Instructions for Use

1. Fix the tissue, deparaffinize routinely to water, and rinse with distilled water for 1–2 min.

2. Place in 3% acetic acid solution for 3 min.

3. Without water rinsing, place in Alcian blue staining solution for 15–30 min.

4. Rinse with distilled water three times, each for 1–2 min.

5. Place in periodic acid solution for oxidation for 5–10 min.

6. Rinse slightly under running water, then rinse with distilled water twice.

7. Immerse in Schiff reagent for 10–20 min.

8. Rinse with sulfite solution three times, each for 2 min.

9. Rinse under running water for 2 min, then rinse with distilled water twice.

10. Place in hematoxylin staining solution for nuclear staining for 1–2 min.

11. Differentiate with acidic differentiation solution for 2–5 s, then rinse thoroughly with water.

12. Rinse under running water until blue color returns.

13. Dehydrate routinely with absolute ethanol. Clear with xylene and mount with neutral balsam.

Precautions

1. Deparaffinization of sections should be as thorough as possible; otherwise, staining results may be affected.

2. Oxidation time with periodic acid should not be too long. The optimal temperature for oxidation is 18–22°C.

3. Alcian blue staining solution, periodic acid solution, Schiff reagent, and hematoxylin solution should be stored tightly sealed at 2–8°C. Avoid excessive exposure to sunlight and air during use. It is best to take them out 30 minutes before use to allow them to return to room temperature, and use them in a dark place away from light.

4. Acidic differentiation solution should be replaced frequently with fresh solution. Differentiation time should be determined based on section thickness, tissue type, and the freshness of the acidic differentiation solution. Additionally, the rinsing time with tap water after differentiation should be sufficient.

5. The action time of sections in periodic acid solution and Schiff reagent is crucial and should be determined based on section thickness, tissue type, etc.

6. When counterstaining cell nuclei with hematoxylin staining solution, staining should be light to avoid affecting the observation of AB-positive substances. The purpose is to prevent cytoplasmic or mucin staining from masking the color of Alcian blue.

7. The staining sequence of the Alcian blue-PAS combined technique can affect the final results. When the PAS technique is performed before Alcian blue staining, neutral mucin and glycogen may stain purple. Conversely, when Alcian blue staining is performed before the PAS technique, these substances can be stained in the expected reddish-purple color.

8. Preparation of sulfite solution: Mix 7.5 mL of 1 mol/L dilute hydrochloric acid, 7.5 mL of 10% sodium metabisulfite, and 130 mL of distilled water.

9. For your safety and health, please wear a lab coat and disposable gloves during operation.

Staining Results

Glycogen, neutral mucin, various glycoproteins	Reddish-purple
Acidic mucin (sulfomucin and sialomucin)	Blue
Proteoglycans and hyaluronic acid	Blue

Note: Cells or tissues containing both neutral and acidic mucin may stain in varying shades from bluish-purple to purple.

Storage and Shipping

Storage

Store at 2-8°C,Protected from light

Shipped In

Wet ice

Stability And Storage

Store at 2-8℃ long term (12 months). Store in the dark.