Questions tagged [fret]
FRET is the abbreviation of Fluorescence resonance energy transfer.
5
questions
1
vote
1
answer
46
views
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 ...
- 11
9
votes
1
answer
378
views
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 ...
- 93
1
vote
1
answer
85
views
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 ...
- 401
3
votes
0
answers
73
views
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 ...
- 4,109
4
votes
1
answer
1k
views
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 ...
- 4,109