Re: X-gal , GMA

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From:rschoonh@sph.unc.edu (by way of histonet)
To:histonet <histonet@magicnet.net>
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Neal,

GMA is (in it's unpolymerized state) miscible with water.  You can, though it's
expensive, use graded (with water) polymer if you decide to go this route let
me know and I'll give you some pointers.

-- Begin original message --

> I am trying to embed and section in JB-4 tissues that have been X-gal
> stained.  I am infecting the mouse mammary gland epithelium with a virus
> that encodes an E. coli b-galactosidase reporter gene.  The main problem
> that I am running into is that the acetone I normally use to dehydrate
> tissues partially extracts the X-gal stain, resulting in a smeared
> appearance in my sections.  I am going to try dehydrating in ethanol or
> even methanol.  What I am really looking for is any protocol that describes
> dehydration of X-gal stained tissues without the extraction of the X-gal
> stain.
>
> Neal Beeman
>
> PhD student
>
> University of Colorado School of Medicine

-- End original message --
regards,
Bob
Robert Schoonhoven
Laboratory of Molecular Carcinogenesis and Mutagenesis
Dept. of Environmental Sciences and Engineering
University of North Carolina
CB#7400
Chapel Hill, NC 27599
Phone
office 919-966-6343
   Lab 919-966-6140
   Fax 919-966-6123

**I'm willing to make the mistakes if someone else is willing to learn from
them**




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