Author Topic: Problems with antigen exposition in retinal cells  (Read 2920 times)

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Offline SCristina

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Problems with antigen exposition in retinal cells
« on: January 24, 2014, 11:38:15 AM »
Our group are working with retinal tissue fixed with 4% PFA. 

We did antigen retreival with citrate sodium ph6 with a pressure cooker during 3 minuts.
The immunostaning was against GFAP (1:200). We also used 10% normal goat serum and 0,5% BSA as a blocking solution.

The problem was that the staining was really low. Any suggestion? Any permeabilization step could be positive? 


Thanks!   :)     

Problems with antigen exposition in retinal cells
« on: January 24, 2014, 11:38:15 AM »

Offline gula

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Re: Problems with antigen exposition in retinal cells
« Reply #1 on: January 25, 2014, 09:35:20 AM »
You can prolong your antigen retrieval. Try serial sections and do HIER with 5 to 15 min.
If your tissue is properly fixed, it should withstand the procedure.
Important: adhesive slides, cooling step after HIER for 10-15 min.

gula

Re: Problems with antigen exposition in retinal cells
« Reply #1 on: January 25, 2014, 09:35:20 AM »