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Dot blot is a straightforward method for detecting biological samples like proteins or nucleic acids. Unlike western and southern blotting, samples are not blotted from a gel but are directly dotted onto a membrane for detection. Due to its simplicity, dot blot supports various applications, including monitoring labeling efficiency, estimating protein concentration, and comparing antibody performance. For example, to determine labeling efficiency with a biotin-labeled sample, one would compare it to a known biotinylated reference standard. This involves making serial dilutions of both the test sample and the reference standard, ideally using a 10-fold dilution for a comprehensive analysis.
