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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