Osteoblast Differentiation Stain (Alizarin Red S)

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Alizarin Rd Staining

  1. Rinse cells very gently one time with water and then very gently add 2% Alizarin Red S stain (preparation below) for 5-10 min at RT
  2. Aspirate stain and wash cells very gently several times with water until wash is pretty clear (can be a very light red).
  3. Add PBS to the well to fix the stain and then aspirate off relatively quickly.  Allow well to air dry completely.
  4. Take phase images.

Preparation of Alizarin Red S

  1. dissolve 2g Alizarin Red S in 100mL dH2O and adjust to pH 4.1-4.3 with ammonium hydroxide
  2. stain can be stored at RT.