beta-gal staining of BHK2

[Harold Drabkin]

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
M>appreciated.


M. Alexeyev:

Here is a procedure that I have used; no staining of untransfected
cells:

Staining cells for b-galactosidase 

Staining solution: 

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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