I often use Chemiluminescent System for western blots. I always make sure the bands are not saturated and then perform densitometry in ImageJ. Recently, I was told that Chemiluminescent System cannot be used for western blot quantification. Is this true? Any insight? Thank you!
If you include standards of purified protein on the blot in order to make a standard curve, then you should be able to get quantitative information by the method you mentioned. Without a standard curve, you should not take the quantitation too seriously because you don't know the relationship between the signal and the amount of antigen.
If you include standards of purified protein on the blot in order to make a standard curve, then you should be able to get quantitative information by the method you mentioned. Without a standard curve, you should not take the quantitation too seriously because you don't know the relationship between the signal and the amount of antigen.
Sorry I could not get what does that mean? So can an imaging system be used for such quantification or densitometer is better for serum protein electrophoresis?
For a gel that has been stained for protein with a colored dye, such as Coomassie Blue, a densitometer is the most quantitative method. An imaging system with a white light transilluminator can also be used as long as the light is uniform over the entire gel.
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