2% Alizarin Red S (mineralisation stain) recipe

Base: 100 mL · pH 4.1–4.3 (adjust with dilute ammonium hydroxide) · Updated 2026-10-01

Stains calcium deposits from osteoblast differentiation red: 2% Alizarin Red S, pH 4.1–4.3.

🧮 Volume calculator
ComponentAmount Final concentration (1X)
Alizarin Red S1 g2% (w/v)
Water (distilled)to final volume

Steps

  1. Dissolve in about 80% of the final volume of water.
  2. Adjust to pH 4.1–4.3 with 0.1–0.5% ammonium hydroxide (pH is critical).
  3. Bring to the final volume with water.
  4. Filter and store dark at 4 °C (about a month).
  5. Stain fixed cells for 20–30 min and wash several times with water.

Good to know

FAQ

Everything is stained red.

Usually the pH is off or washing was insufficient — recheck pH 4.2 and wash more.

This is a widely used standard composition. Amounts depend on the hydrate form and purity of your reagents — check the formula weight (FW) on the bottle and follow your institution's safety rules and the manufacturers' instructions.