Description Colloidal ferric ions are absorbed principally by carboxylated and sulfated mucosubstances at low pH. The excess reagent is washed out and the classic Prussian blue reaction (iron stain) is used to demonstrate iron bound to the tissue. Colloidal Iron Stain Kit is designed to demonstrate carboxylated and sulfated mucopolysaccharides and glycoproteins in the tissue. It is suitable for any well fixed paraffin embedded tissue. Kit Components SKU# Reagent Size (ml) VB-3027-1 Acetic Acid Solution (12%) 500 VB-3027-2 Hydrochloric Acid Solution (3%) 125 VB-3027-3 Potassium Ferrocyanide Solution (3%) 125 VB-3027-4 Colloidal Iron Stock Solution 125 VB-3027-5 Van Gieson’s Solution 125 Storage Store at room temperature Protocol Thing to do before staining Working Colloidal Iron Solution. Acetic Acid Solution (12%) —————5 ml Distilled Water ——————————-15 ml Colloidal Iron Stock Solution———– 20 ml Mix well. Working Iron Stain Solution Hydrochloric Acid Solution (3%) ——–20 ml Potassium Ferrocyanide Solution——- 20 ml Mix well Procedures Deparaffinize in xylene I for 6 minutes and II for 6 minutes. Rehydrate Ethanol 100% (1 minutes) Ethanol 100% (1 minutes) Ethanol 95% (1 minutes) Ethanol 95% (1 minutes) Ethanol 70% (1 minutes) Rinse in DI water (2 minutes). Cover tissue section with Acetic Acid Solution (12%) for 30 seconds. Place slide in Working Colloidal Iron Solution for 30 minutes with occasional gentle agitation. Use once and discard. Rinse thoroughly in 3 changes of Acetic Acid Solution (12%) for 2 minutes each. Stain slide in Working Iron Stain Solution for 10 minutes with occasional gentle agitation. Use once and discard. Rinse in 3 changes of distilled water. Stain tissue section with Van Gieson’s Solution for 30 seconds. Dehydrate in 3 changes of absolute alcohol. Clear with 3 changes of xylene (5 minute per change) Mount coverslip onto glass slide with Permount or some other suitable organic mounting medium. Expected Results: Acid mucopolysaccharides and sialomucins——–deep blue Collagen———————————————————red Control Tissue: Skin, Small intestine or Colon. 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) VB-3027 User Manual VB-3027 MSDS
