b-Galactosidase Color Assay for Cultured Cells

b-Galactosidase Color Assay for Cultured Cells


1. For transient expression assays, wait until 2 days after
the transfection to begin the color assay.  Aspirate the media and
rinse the cells twice with PBS.

2. To fix the cells, add 5 ml of a 0.5% Glutaraldehyde in
PBS (pH 7.2) solution; incubate @ room temp. for 10 minutes.

3. Rinse the cells twice with PBS.

4. Stain the cells in [5 mM Potassium Ferricyanide, 5 mM
Potassium Ferrocyanide, 2 mM MgCl2, and 1 mg/ml X-gal] overnight in
the dark @ room temp.


Notes: The 25% Glutaraldehyde stock solution is stored @ -20o C.

200 mM stocks of the potassium solutions are prepared in PBS and
stored in the dark @ 4o C.

0.01% NaDOC and 0.02% NP-40 may be included in the staining
solution, but they are not essential.


From the Laboratory of Dr. Allan Bradley 
Baylor College of Medicine, Houston, Texas