FRET or BRET between DsRed and GFP or Renilla luciferase ?

Janos, Szollosi szollo at jaguar.dote.hu
Tue Aug 8 11:56:48 EST 2000


Hi Holger,

I have not experience yet with the DsRed protein, but I checked the spectra 
and I was also told by the Clonetech, that this dye not a perfect acceptor.

The major problem is that it has long excitation spectra, it can be excited 
quite well at the excitation wavelength of the donor (e.g. YFP, GFP) 
therefore the enhancement is not too good.

This means that you will have too much direct signal and only a small 
enhancement due to the FRET.  Your FRET signal could be in the error range 
of your intensity measurement.

The Clonetech would like to improve the excitation spectrum of the 
DsRED.  I would wait for that!

I hope this helps

Regards

Janos


At 12:08 PM 8/3/00 +0100, you wrote:
>Hi,
>
>I wanted to measure protein-protein interactions using Clontech´s DsRed in
>FRET- or BRET-assays, but havn´t see much effects so far. So my question is:
>
>Had anyone seen significant FRET between GFP and the Red-FP or BRET between
>Renilla luciferase and RFP ?
>
>I didn´t see much effects even with a GFP-RFP fusion protein, but I can not
>rule out that this might be a specific problem of my construct.
>
>So if anyone has positive or negativ proof for FRET or BRET using RFP,
>please send a comment.
>
>
>Sincerly,
>
>Holger Fehr
>
>
>---------------------------------------------------
>
>Dr. Holger Fehr
>Inst. Physiological Chemistry II
>University of Wuerzburg
>Germany
>
>fehr at biozentrum.uni-wuerzburg.de
>
>
>
>---

Janos Szollosi
University of Debrecen
Medical and Health Science Center
Faculty of Medicine
Department of Biophysics and Cell Biology
P.O.Box 39, Nagyerdei krt. 98
H-4012 Debrecen
Hungary
Phone/Fax:  (36) (52) 412-623
E-mail:  szollo at jaguar.dote.hu


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