Re: BrdU - the never ending story....

From: Mark Shlomchik (mark.shlomchik@yale.edu)
Date: Thu Sep 30 1999 - 17:22:13 EST


At 7:44 PM +0200 9/29/99, Christoph Goettlinger wrote:
>I have a question concerning a MoAb to BrdU, which is designated BU-1.
>Right now I am trying to stain for BrdU as well as for different cell
>surface markers. So, not all methods are appropiate. Looking for an
>resolution, I found an old dicussion about that topic in the archive of
>this forum (dated back to 1995). It was said that with the MoAb BU-1 you do
>neither need to fix the cells nor do you need to denature the DNA. I was
>really happy to find that messages. But following up the discussion was
>desillusionating when I tried to find this clone.
>I would be very happy if somebody could provide me with the information
>where I can get this clone BU-1 from. Any information is more than welcome.
>Thank you very much.
>
>My work address is: 	Christoph Goettlinger
>			University of Cologne
>			Weyertal 121
>			Germany
>
>			email: c.goettlinger@uni-koeln.de

We are currently doing alot of BrdU staining and have similar goals 
and in fact would also like to do DNA/RNA analysis of cells post BrdU 
staining.  However, the only Abs we have gotten to work are those 
from BD and Phoenix Flow, both of which are equivalent.  We however, 
need to do fixation/perm as well as DNAse tx.  We have the best 
results using EtoH as per the original Tough and Sprent paper.  It 
seems this part may be optional if the cells are kept fix/perm long 
enough(?)  I am not aware of any other clones that even work in this 
system (the Caltag Ab did not work here nor in immunohistochem).

Another issue is the inordinate expense of the BrdU Abs. It would 
seem that the BD Ab, at $400 per 50ug (i.e at $8,000/mg) is the 
world's most expensive Ab.  It would be so nice to get hold of a 
clone that I could grow in the lab. Anyone who could help with this 
would be most appreciated.

Cristoph, if you get any private replies I would greatly appreciate 
if you could forward them to me.

Mark Shlomchik, MD, PhD
Associate Professor of Laboratory Medicine
and Immunobiology
Yale University School of Medicine

203-688-2089
203-688-2748 (fax)
mark.shlomchik@yale.edu



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