Crystals during X-gal staining

Martin Leach leach at mbcrr.harvard.edu
Thu Apr 21 11:16:52 EST 1994


In article <CoMBtv.I1M at freenet.carleton.ca>, ar229 at FreeNet.Carleton.CA
(Allison Haggarty) wrote:

> 
> 
> I have been using X-gal to stain cells blue after transient tranfections. 
> The cells are blue allright but there are lots of crystals in the stain so
> the results are not very photogenic.  The problem seems to arise when the
> X-gal stock (10% in DMF, frozen at -20 C) is diluted 1:100 in the working
> buffer for staining (5mM K ferricynanide, 5mM K ferrocyanide, 2 mM MgCl2,
> made up in PBS).  As soon as the X-gal hits the aqueous buffer the
> solution goes cloudy. 
> Does anyone know how to prevent this? Am I stuck with filtering? 
> Thanks.
>

Warm it to 37C for 10-20mins then filter it

Otherwise u get crystals forming when u place it on the cultures.

Martin.
-- 

.....          Martin Leach                Email:leach at mbcrr.harvard.edu 
   _|____      Dept. of Pharmacology       Phone: (617) 638-5323        
   / o  /      Boston Univ. School of Med. Fax:   (617) 638-4329         
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