Re: LacZ

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From:Jamie Erickson <JErickson@genetics.com>
To:HistoNet@pathology.swmed.edu
Reply-To:
Date:Tue, 31 Aug 1999 17:29:24 -0400
Content-Type:text/plain

Here is Our Protocol for staining for LacZ, the enzyme method not the antibody method.

For 100ml     100ml PBS NP-40
                     200 ul 1M mgCl2
                     800 ul ferricyanide
                     800 ul ferrocyanide
 Filter then Add:
               2000ul X-Gal (100mg/2000 ul DMF (dimethyl formamide)

Gultaraldehyde
                      0.05% GTA

1mM MgCl2
                      0.09521g/Liter

1.  PBS for two minutes 
2. Fix twice in PBS/1mM Mg/CL2 for  10 minutes each.
3. Incubate in X-Gal Overnight.
4. Rinse in running water for 1 hour
5. Counter stain in Nuclear Fast Red For 10 minutes.
6. Rinse in dH2O).
7. Coverslip w/ crystal mount (biomedia)
8. Put Slides in oven @ 70 degrees C for 30 minutes.

Jamie
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