Subject: beta-gal staining of BHK21 cells
M>I am staining BHK21 cells transfected with plasmid encoding
M>beta-galactozidase. As a result I can see blue cells in both negative
M>control and in my experiment (the numbers are almost equal). My solutions
M>are made in PBS (pH7.3). Any remedies/ideas-suggestions will be
Here is a procedure that I have used; no staining of untransfected
Staining cells for b-galactosidase
5mM potassium ferricyanide (165 mg/100ml), 5mM potassium ferro-cyanide (210 mg /100ml), 2 mM MgCl2 (0.2ml of 1M stock in 100
Make the above up in 100ml with PBS, and store at -20oC.
1. Fix cells for 10 minutes in a 1:20 dilutiion of 2.5% glutaraldehyde (dilute with PBS).
2. For staining place 315 ml of 40 mg/ml X-gal (in DMF) and per 9 ml of stain per 10cm dish. Prewarm thissh. Place stain on
3. Remove the staining solution, and overlayer cells with 50% glycerol. Examine under the microscope.
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