e.g. I have a set of GFP-based membrane proteins, I detect the fluorescence intensity. I could say the more intense detected fluorescence, the more the amount of fusion protein. But what is the accurate relation between both of them.
e.g. linear? log? exponent？ or else？
Do you have some recommended papers to say the question?
In general, fluorescence intensity is directly proportional to the concentration of fluorophore, as long as (1) the fluorophore concentration is not so high that its absorbance becomes significant (inner filter effect), or (2) the fluorescence intensity is so high that the detector becomes saturated.
You have GFP labeled membrane proteins.
Why is it important to know the concentrations of these proteins based on fluorescence? You can easily do a western and probing it with anti-GFP Abs to see the relative conc. of your proteins.
Are you dealing with purified proteins, or with cells expressing the proteins viewed by microscopy or flow cytometry? With microscopy and flow cytometry, the pathlength through the sample is so short that the inner filter effect is not an issue.
Instrument software will generally tell you if the detector is saturated by excessive fluorescence intensity.
If you are working with purified proteins, use a protein assay to measure the amount of protein.
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