RE: Borax pretreatment for brdU

From:Chris van der Loos

Dear Carl,
I sure realize that we should have done so, but we haven't. It was once 
tested successfully by somebody who was in a hurry and did not like to 
prepare fresh formamide- and HCl reagents (at that time a 
formamide-treatment at 60C was still included, no borax). Sorry, but that's 
the way it sometimes goes! Because his aim was just qualitatively we never 
compared the two different methods, although we should have done it. Since 
we are no regular BrdU-appliers it was never done after.
As you indicated a difference in positive nuclear numbers, I totally agree 
with you that a comparison needs to done.
Chris van der Loos
Academic Medical Center
Dept. of Cardiovascular Pathology
Amsterdam, Netherlands

Carl wrote:
 >Date: 5 Feb 2003 03:30:28 -0600
 >From: Carl Hobbs 
 >Subject: Borax pretreatment for brdU
 >"Dear David,
 >Considering the the use of HCl etc. you obtained an older BrdU IHC staining
 >protocol. Here we once tried to stain BrdU containing FFPE sections just by
 >using a standard antigen retrieval method (citrate pH6.0). It worked
 >perfectly fine!"
 >Chris, Interesting! I tried this last week and, altho' I got positive
 >nuclei, the number of positives was far higher when I used the traditional
 >HCL/ trypsin method. Did you compare the two?
 >Carl Hobbs
 >Laboratory Manager
 >Molecular Neurobiology Group
 >MRC Centre For Developmental Neurobiology
 >New Hunts House
 >Guys Campus
 >kings College London
 >SE1 1UL 

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