Do you know that journals like the JBC strongly caution against the use of housekeeping proteins for normalization? Most journals now demand rigorous standards for presentation of western blotting data. While housekeeping proteins (HKP) have traditionally been used for decades, the problems with their usage have come to the fore only recently. More validation is required for presenting data using HKP. Normalization using total proteins present in the sample has gained significant traction and has proved to be a reliable method for normalization of western blots. Here we illustrate why the total protein normalization (TPN) method is superior and the way to go for western blotting.
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