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Dot blot is a straightforward technique for detecting biological samples, such as proteins and nucleic acids. Unlike western or southern blotting, the sample is applied directly onto a membrane, without prior gel transfer, allowing for quick detection. Due to its simplicity, dot blot is applicable in various settings, including monitoring labeling efficiency, estimating protein concentration, and comparing antibody performance. For instance, to assess labeling efficiency of 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 reference standard, ensuring identical dilution methods, with a 10-fold dilution serving as a suitable starting point for a broad range of samples.