Alcian Blue, also known as Alician Blue or Alcyan Blue, is a copper - phthalocyanine conjugated dye originally used for textile fiber dyeing. As a cationic dye, it binds to acidic groups, namely, it forms insoluble complexes with anionic groups such as carboxyl groups and sulfate groups contained in tissues. Alcian Blue is composed of a central copper - containing phthalocyanine ring linked to four isothiourea groups via thioether bonds. The isothiourea groups are moderately basic, endowing Alcian Blue with cationic properties. At pH 2.5, carboxyl groups in tissues ionize and carry a negative charge, forming salt bonds with the cations in Alcian Blue, thus staining acidic mucous substances (sulfomucins and sialomucins) with carboxyl groups.
Alcian Blue Staining Solution (pH 2.5) is also called standard Alcian Blue Staining Solution. Different pH values of the staining solution can be used to distinguish the types of mucous substances. At pH 1, carboxyl groups (COOH) are not stained while sulfate groups (OSO₃H) are stained. At pH 2.5, carboxyl groups are well - stained but sulfated mucins are poorly stained, resulting in the staining of sulfomucins and sialomucins. Neutral mucins (such as those in gastric mucosa and Brunner's glands) do not react with Alcian Blue. This reagent is only applicable to scientific research and not for clinical diagnosis or other purposes.
A774858
| Component | 3×50 mL | 3×100 mL | Storage |
|---|
| A774858A | Alcian Blue Acidification Solution | 50 mL | 100 mL | RT. | | A774858B | Alcian Blue Staining Solution | 50 mL | 100 mL | RT. Store in the dark | | A774858C | Nuclear Fast Red Staining Solution | 50 mL | 100 mL | RT. Store in the dark |
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Materials to Be Prepared by Users
1. Graded ethanol solutions
2. Distilled water
Operating Procedures (for reference only)
1. Deparaffinize the sections with xylene, rehydrate them through graded ethanol solutions, and then transfer them into distilled water.
2. Immerse the sections in Alcian Blue Acidification Solution for 5 minutes.
3. Stain the sections in Alcian Blue Staining Solution for 30 minutes.
4. Rinse the sections with running water.
5. Counterstain the sections in Nuclear Fast Red Staining Solution for 5-10 minutes, followed by rinsing with running water.
6. Dehydrate the sections with graded ethanol solutions, clear them with xylene, and mount the sections with neutral balsam.
Staining Results
Acidic mucins (sulfomucins and sialomucins): Blue
Proteoglycans and hyaluronic acid: Blue
Cell nuclei: Red
Precautions
1. Use 10% neutral formalin as the fixative.
2. For the selective identification of sulfomucins and proteoglycans, Alcian Blue with a low pH value (pH=1.0) should be used, i.e., adjust the pH value to 1.0. The staining procedure is the same as that for Alcian Blue at pH=2.5, but the staining time should be appropriately extended.
3. Opened reagents should be used up within 6 months after opening. Tighten the bottle caps immediately after each use to avoid volatilization or deterioration.
4. For your safety and health, wear a lab coat and disposable gloves during operation.
5. This product is for research use only and shall not be used for other purposes.
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