Housekeeping proteins Western blot immunoblotting loading control total protein staining. What is the best nuclear protein loading control (western blot) Hi all, I am doing some methylation work and need to report nuclear protein expression changes responsible for the. I conclude that TPS should be the preferred loading control in future Western blot approaches. Consequently, this review article offers an orientation in selecting the appropriate staining type. Nine benefits of TPS are discussed and seven different variants are critically evaluated by comparing technical details. The objective of this review article is to collect and compare information about TPS options and to invite users to reconsider their applied loading control. Although these and further significant advantages have been demonstrated over the past 10 years, only a small percentage of laboratories take advantage of it. Further, the broad dynamic range of TPS solves the issue of HKPs that commonly fail to show loading differences above small loading amounts of 0.5-10 μg. Total protein staining (TPS) represents the actual loading amount more accurately than HKPs due to minor technical and biological variation. Staining of the total protein on the blotting membrane has emerged as a better loading control. However, this loading control has multiple drawbacks. Currently, housekeeping proteins (HKPs) like actin, tubulin, or GAPDH are often used to check for equal loading or to compensate potential loading differences. This review article focuses on a significant, but not yet well-established, improvement concerning the internal loading control as a prerequisite to accurately quantifying Western blots. ![]() Western blots are continuously applied and developed by the protein community. Western blotting is an analytical method widely used for detecting and (semi-)quantifying specific proteins in given samples.
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