When doing fluorescence intensity measurements, do you need another fluorescent protein to normalise it with?

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I want to perform intensity analysis for a protein that is expressed on the nuclear lamina. I would like to identify what is the average expression of the protein in the cells at different time points.

Do I need to normalise such value for another marker that is known to be unchanged in the cell or is this not needed in IF?

Sung-Jun Lee

It depends on the wavelength (nm) range of your fluorescence of interest. If applicable in your study, well-known control proteins such as GFP, eGFP, or eYFP can be used. I agree with the idea that control markers are necessary to normalize the fluorescence signals, especially for your case.

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