In a soon to be published paper that will appear in Analytical Biochemistry, a team of scientists from the LMU and Federal Office for Radiation Protection in Germany and Bio-Rad Laboratories compared two methods of protein normalisation utilized in western blot analysis. In the first method, known as housekeeping protein (hkp) normalisation, the signal of the protein of interest was divided by the signal of a housekeeping protein such as GAPDH. In the second method, western blotting signal from the protein of interest was normalised by the total amount of protein (from the same lane as the protein of interest) as quantified by stain-free technology.
The authors found that Stain-Free technology appears to be more reliable, robust and more sensitive to small effects of protein regulation when compared to HKP normalisation. Stain-Free technology offers the additional advantages of providing check points throughout the western blotting process by allowing rapid visualization of gel separation and protein transfer.
For more information download Stain-Free Technology as Normalisation Tool in Western Blot Analysis.