Is Reuse of Electrophoresis Buffer in Western Blotting Acceptable?
In most cases, reusing electrophoresis buffer in Western blot experiments is not recommended. The buffer components—such as Tris, SDS, and other ionic species—may become depleted or accumulate contaminants, including residual proteins, cellular debris, and other impurities introduced during electrophoresis. These changes can compromise the consistency and accuracy of protein migration and ultimately affect the reliability of experimental outcomes.
Furthermore, previously used running buffer carries a risk of microbial or cross-sample contamination, which may introduce artifacts or variability in subsequent experiments. To maintain experimental reproducibility and data integrity, it is advisable to prepare and use fresh electrophoresis buffer for each Western blot run.
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