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which normalization strategy is best for your experiment well it depends you can use housekeeping proteins for normalization as long as you validate that their expression is not changing with treatments or experimental conditions or you can choose an easier more straightforward method called the total protein staining method in the instructions to authors the journal biological chemistry states normalization of signal intensity to total protein loading assessed by staining membranes using Kumasi blue path OS or other protein stains is preferred total protein detection is becoming the gold standard for normalization of protein loading after transfer but prior to immuno detection the membrane is treated with a total protein stain to assess actual sample loading across the blot because this internal loading uses the combined signal from many different sample proteins in each lane error and variability are minimized this antibody independent method corrects for variation in both sample co
