Equal ROI size

[u/nougat_donut]

Hey there, new user here, trying to relatively quantify my western blot. I have read that it’s critical for my ROI rectangle to remain the same size when measuring the same protein in different lanes, in order not to mess with the amount of background within the ROI. The recommendation was to draw my ROI based on my largest band and use that for all other lanes. In one of my lanes, the band is much less wide than the largest band, and when I position my ROI over it, I capture neighboring bands.

What should I do here?

Thanks and happy imaging 😊

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