Microplate cell growth problem

Wolfgang Schechinger wgschech at med.uni-tuebingen.de
Mon Sep 28 07:17:00 EST 1998


Hi Morton!

I'ts probably the naturally occuring isotope mixture of Br. BrdU is 
*not* radioactive; that's the reason why we are using it. The assay 
is colorimetric: BrdU is incorporated into DNA instead of 
Thymidine. BrdU then is recocgnized by an antibody, this antibody is 
labelled with peroxidase, peroxidase cleaves superoxide wich then  
turns a chemical into a blue dye.

But thanks for the hint on 14C doses.

Wolfgang

> Just a thought:
> 
> What Br-isotope are you using? Outer cells will recieve less
> crossfire radiation from the surrounding wells and therefore might
> be less growth inhibited than the center cells.
> 
> I don't know what doses you use, but I have the experience with
> hamster fibroblasts that even 1000 Bq/mL (C-14 thymidine) makes a
> detectable decrease in growth rate.
> 
> Morten Lindow
> 
> 
> 
> 
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Wolfgang Schechinger         
University of Tuebingen, Germany
email: wgschech at med.uni-tuebingen.de * wwWait: http://www.medizin.uni-tuebingen.de/~wgschech/start.htm
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