Questions tagged [fret]

FRET is the abbreviation of Fluorescence resonance energy transfer.

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Method to detect Protein aggregation

I have a specific human protein that was purified from E. coli. This protein was tagged with MBP and diluted in an elution buffer. This protein has a tendency to aggregate, and when it does, it can ...
Sketsawy's user avatar
9 votes
1 answer

Why is there a λ⁴ in the spectral overlap integral in FRET calculations

The Förster resonance energy transfer (FRET) spectral overlap integral looks like: $$ J=\int_{0}^{\infty}\bar{F}_{D}(λ)ε_{A}(λ)λ^4 dλ $$ Where $λ$ is the wavelength, $\bar{F}_{D}(λ)$ is the normalized ...
Weiwei's user avatar
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Why such strange enzyme kinetics?

I measured some enzyme kinetics in a practical course using a substrate-based FRET assay. Unfortunately some of my plots show weird effects. There was always a decrease in signal after 35 minutes. But ...
Mourinho_1's user avatar
3 votes
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Removal of the Initial Methionine in Venus for FRET

I'm working on building some FRET reporters. In addition to a cleavage site (of varying composition from 15-18AA), a 1 AA linker, I'm using Venus and Cerulean. Initially I was worried that 18AA ...
Atl LED's user avatar
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4 votes
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Is there an optimal composition and length for protein linkers in FRET?

I'm designing a protein that I'd like to use in FRET reporting. General idea on the shape is: FRETprotein1--Linker--CleavageSite--Linker--FRETprotein2. I would like to know what AA are best for the ...
Atl LED's user avatar
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