Fluorescence tutorial sought

John H. Collins collins at UMBI.UMD.EDU
Thu Mar 18 06:50:13 EST 1999


Does anyone know of a good, online, basic text for this sort of 
thing, i.e. fluorescence spectroscopy and its application to proteins?
-John Collins, Medical Biotechnology Center, Baltimore

On 18 Mar 1999, Bhupesh Taneja wrote:
> Hi all!
> 1)      I want to do folding-unfolding studies with my protein.
> Unfortuantely, the single tryptophan is partially exposed. (as from
> homologous xtal strucures and by quenching with KI). The emission maxima
> I obtain is at 346 nm for the native protein with a broad band width and
> for fully unfolded protein(thermal) at 350 nm. Is it possible that most
> of the protein molecles are in an unfolded state? How do I check this?
> (The protein is in 10mM Tris-Cl, pH 7.5).  From other structures ~25-30%
> of trp is exposed.  Also KI would bind to exposed Trp in folded and in
> unfolded proteins.
> 2)      How is the correction for decrease in fluorescence due to increasing
> temperature is done in thermal unfolding?
>                 Thanks for all suggestions.
>                                         Bhupesh. 17-3-99.
>                                 bhupesh at lion.imtech.ernet.in
>                                 bhupesh at bragg.imtech.ernet.in
> 
> 



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