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Dot blot is a straightforward method for detecting biological samples such as proteins or nucleic acids. Unlike western and southern blotting, samples are applied directly onto a membrane rather than being transferred from a gel. Due to its simplicity, dot blot has diverse applications including monitoring labeling efficiency, estimating protein concentrations, and comparing different antibodies. For instance, to assess labeling efficiency with a biotin-labeled sample, one would compare it to a known biotinylated reference standard. This involves creating serial dilutions of both the test sample and the reference standard in the same manner, with a 10-fold serial dilution being a recommended starting point for obtaining a broad sample range.