Thinking about doing FRET - please point me in the right direction

Dima Klenchin klenchin at REMOVE_TO_REPLY.facstaff.wisc.edu
Tue Jan 23 17:23:46 EST 2001


Hello, 

I am thinking that a protein I study might self associate
under some specific conditions (binding to cell organells).
I have a lot of the protein available and I was thinking if
doing FRET is a good way to assay for this kind of thing.

The idea is to covalently label two pools of protein with two
different dyes, donor and acceptor, and then see if and when
there is an increased fluorescence appearing. 

Problem is, I don't know much about the technique and thus
far can hardly make an educated choice of what dyes to use,
nor I can sense any caveats in using this approach. 

I just started to dig through the literature and so far results
are chaotic. 

Any comments will be greatly apprecaited. Is this feasible
at all? Where to start? Specific books, papers I must read?
Any other advice?

Thank you.

        - Dima
 






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