[Histonet] Re: Fluorescent DAB?????

[AGrobe2555 <@t> aol.com]

Gayle Callis wrote: "DAB will fluoresce when excited at same wavelength as  
FITC."
 
My questions are:  Assuming that all the parameters were kept  equivalent 
within a "batch" of slides:  
 
Can it be used in image analysis to quantify expression of a particular  
epitope of interest among different samples in a batch of stained slides?  
 
Or is DAB precipitate more of a random quantity depending on substrate  
availability/enzyme activity and therefore cannot be used to compare among  slides 
(I realize that enzyme kinetics make the word "random" a bit of  an 
contradiction in this sense)?  This is based more upon the premise  that the fluorescent 
secondary only has so many places to bind vs. HRP that  creates more color 
the longer it works......(at least until the reaction is  stopped or the 
substrate/cofactors are exhausted).  
 
Thanks,
Albert  

Albert C.  Grobe, PhD
International Heart Institute of Montana Foundation
Tissue  Engineering Lab, Saint Patrick  Hospital





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