Dear Joe,
               Interesting..!! Have you found an increase in background 
fluorescence proportional to the time used for heat based Ag retrieval 
in old paraffin embedded tissue? Can microwave and pressure cooking can 
be used for all antigens or does it vary for Ag to Ag? Does any 
histonetter have any experience with fluorescent IHC of telomerase using 
microwave techQ?

Thanks
Swaram


Galbraith, Joe wrote:

>Amy:
>
>We do our blocking prior to retrieval.  We've had similar discussions in the lab in the past and our experiments have failed to demonstrate any large difference, whether blocking is before or after retrieval.  We have not detected any return of background that had been previously blocked as a result of the heat retrieval approaches that we routinely use.  Obviously some experimentation using your own methods would be best as the intensity and conditions of retrieval techniques do vary.  We keep our heat retrieval approaches standardized but also minimized - just enough to do the trick - which means that our approach tends to be less intense than some labs but our staining still strong and clean.
>
>Good luck
>
>Joe Galbraith
>
>-----Original Message-----
>From: histonet-bounces@lists.utsouthwestern.edu
>[mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Amy
>Porter
>Sent: Thursday, July 15, 2004 9:46 AM
>To: Histonet
>Subject: [Histonet] endogenous alk phos block and HIER
>
>
>Just curious to know what everyone thinks about pretreatments for endogenous products and Heat induced retrieval methods.  Do you do the blocking before the retrieval or after?  I am always worried about effecting the pH treatment from the retrieval if i pretreat for endogenous after retrieval.  On the other hand if I block first and then retrieve, am i re-opening what i just tried to get rid of while retrieving.  I hope this makes sense to someone and opinions would be greatly appreciated.  I know i will get lots of good information, as always - Thanks in advance for all the help.
>
>
>Amy S.Porter, HT(ASCP) 
>Michigan State University 
>Department of Physiology 
>Division of Human Pathology 
>College of Human Medicine 
>portera203@yahoo.com
>
>
>		
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