Description
The Alcian Blue-PAS Stain Kit utilizes the properties of both the Periodic Acid Solution (PAS) and Alcian Blue methods to demonstrate the full complement of tissue proteoglycans. The rationale of the technique is that by first staining all the acidic mucins with Alcian Blue, those acidic mucins which are also PAS positive will be chemically blocked and will not react further during the technique. Those neutral mucins which are solely PAS positive will subsequently be demonstrated in a contrasting manner. The combined result demonstrates both acidic and neutral mucins as well as mixtures of acidic and neutral mucins.
Kit Contents
- VB-3005-1 Alcian Blue Solution——————250 ml
- VB-3005-2 Schiff Reagent ————————–250 ml
- VB-3005-3 Periodic Acid Solution—————250 ml
- VB-3005-4 Mayer’s Hematoxylin Solution—250 ml
Storage Condition:
Schiff Reagent stored at 2-8°C and the others store at room temperature.
Protocol
- Deparaffinize in Xylene I for 6 minutes and II for 6 minutes.
- Rehydrate: ethanol 100% (2 minutes)x2; ethanol 95% (2 minutes)x2; ethanol 70% (2 minutes)x1; Rinse in distilled water (5 minutes).
- Stain with Alcian blue solution for 15 mins.
- Wash well in running tap water for 2 mins.
- Rinse in distilled water.
- Treat with periodic acid solution for 5 mins.
- Rinse in distilled water.
- Stain with Schiff’s reagent for 10 mins.
- Rinse in running tap water for 5 mins.
- Counterstain in Mayer’s hematoxylin for 1 minute.
- Wash in tap water for 5 minutes.
- Dehydrate and Clear with 3 changes of xylene (5 minute per change).
- Mount coverslip onto glass slide with Permount or some other suitable organic mounting medium.
Results
- acidic mucins ……………………………… blue
- neutral mucins ……………………………. magenta
- mixtures of above ……………………….. blue/purple
- nuclei ……………………………………… deep blue
Technical Points
Diastase pre-treatment may be used to eliminate the presence of glycogen, prior to staining in alcian blue.
Note: This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.
Precautions: Handle with care. Avoid contact with eyes, skin and clothing. Do not ingest. Wear gloves.
User Manual and Material Safety Data Sheet (MSDS) (PDF)
