Re: [Histonet] re antigen retrieval

From:"Steven E. Slap"

Hi Carl & HistoNetters

Good point, Carl-  makes you wonder, doesn't it?  According to Dr. 
Shi, the factors that really matter are time, temperature and pH. 
Some antigens turn out to be "easy", others "hard".  It doesn't seem 
to matter too much what solution is used so much, as you say, so long 
as the pH is right for the particular Ab, and the time and 
temperature are accurately controlled.  That being said, I have 
usually found that high pH solutions damage morphology, and prefer to 
use pH 6.0 citrate buffer whenever I can get away with it, and raise 
the temperature or lengthen the time rather than raise the pH to get 
the "hard" aBs to work.

best regards,
Steven

At 9:25 PM +0100 4/19/04, Carl wrote:
>Blew the cobwebs there, John K. I remember reeling with horror at 
>the thought of using it ( I did....once). Was I justified in my 
>fears? It didn't do anything for my MIB1s/p53s , I have to 
>disclose.Neither did Urea........
>NB Must admit, Urea never did anything for me with other Ags( except 
>strip my sections off).
>Also,how come so many of us have best results with different high 
>temp AR solutions, yet with same( it seems) Ab reagents?


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