Alizarin Red Staining for Mineralising Cultures

1. Prepare 2% w/v Alizarin Red S solution by dissolving 500mg Alizarin Red S powder (Sigma, A5533) in 25ml deionised water (d.water) and adjust to pH 4.2 with 0.5% ammonium hydroxide 2. Fix cells in 70% v/v ethanol for 60 min on ice. 3. Wash 3x in d.water and stain with 2% Alizarin Red S solution for 10 minutes at r.t. 4. Rinse cultures with d.water

Alizarin Red staining of human osteoblasts after 14 days culture in medium containing 0.2M hydrocortisone