normalisation of western blot from kodak film - (Oct/13/2011 )
I developed my western blot with ECL PLUS using kodak film, then I need to normalise my protein with actin. I scanned the film and qantify qith Image J, I need to know if my quantification should consider the result on the film or whether I should set parameters of resolution of scanning the film . to have the real proflie of protein level!
which resolution or... I should use?
Ideally you need to use the maximum possible file size/resolution - to eliminate pixel artifacts. You also need to make sure the files are not "lossy" formats (where pixels of similar or the same intensity are grouped and marked as one pixel) such as JPEG/JPG. TIFF/TIF is what you should be using.