B-gal staining on fixed samples? - (Jan/05/2005 )

Does anyone know if you can stain for beta-gal on formalin-fixed, parafin-embedded tissue slices? Most of the protocols I have seen ask for frozen samples, but we have brain sections already prepped in parafin we would like to stain. Any protocols would be helpful, too.

Anyone?? Anything would help....

Here are a few tips.....

I fixed my tissue in:

2.7mL 37% formaldehyde
400uL 25% glutaraldehyde
brought it up to 50mL in PBS, pH 7.6

My stain was:
25mg X-Gal dissolved in 625uL DMSO. Set aside
Dissolve 41mg potassium ferricyanide & 53mg potassium ferrocyanide in 24mL PBS, pH 7.6. Vortex
To this add X-Gal from #1 & 50uL MgCl2
Filter with a syringe filter
Only store for 2 weeks.

I usually stained my tissue first and then used a processing procedure that cuts the time for each step in half. From there I was able to section without a problem.

I hope this helps and gives you some thought at least.

Rachael