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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 directly dotted onto a membrane, avoiding the gel transfer step. This simplicity allows dot blot to support various applications, including monitoring labeling efficiency with probes, estimating protein concentration, and comparing different antibodies. For example, to assess labeling efficiency with a biotin-labeled sample, one can compare it to a known biotinylated reference standard. This involves creating serial dilutions of both the test sample and the reference standard, maintaining identical dilution methods, with a 10-fold dilution recommended for a broad sampling range.
