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Star Republic: Guide for Biologists

X-gal staining of cells

1. Remove cell media, wash cells twice with PBS.

2. Fix cells in PBS/0.05% Glutaraldehyde. Incubate at room temperature for 5 minutes.

4. Remove PBS/0.05% Glutaraldehyde and wash twice with PBS.

5. Stain with PBS containing 1 mM MgCl2, 5 mM K4[FeII(CN)6], 5 mM K3[FeIII(CN)6], plus 1 mg/ml X-gal. X-gal staining solution should be warmed to 37 C for 5-10 minutes before use.

6. Incubate at room temperature or 37 C in dark until desired level of staining is observed (several hours to overnight). Stop staining by washing several times with PBS.