BrDU proliferation assay

[arveiler]

In article <1995Feb23.173459.7706 at ucsvc.ucs.unimelb.edu.au>,
chris at austin.unimelb.edu.au wrote:

> Has anyone had experience with the BrDU labelling assay
> for cell proliferation?  I have attempted to use it in 
> a manner analagous to the continuous labelling of cells
> with tritiated thymidine.  This a simple method giving
> much information of cell cycle time etc.  However, 
> rather than getting a simple increase in labelling over
> time with an eventual plateau when all cells are 
> labelled, I obtained a sine curve with two peaks, the 
> second less than the first.  I would be glad of any 
> explanations (or even half convincing fictions!).
> 
> Thanks in advance,
> Chris Bradley


BrdU may be toxic for some cells; thus in the first time you kill the
sensible one, and you observe a decrease of signal, then you have an
increasing incorporation in a fewer number of cell (smaller peak).
To get a fait idea about the sensibility of your cells just number them
during the BrdU treatment using the blue trypan to check viability

Avoid benveniste-like sine curves in the future!

and good luck...

vekris.u-bordeaux2.fr