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Dot blot is a straightforward method for detecting biological samples like proteins or nucleic acids, paralleling western and southern blotting techniques, but without the need for gel transfer. Instead, samples are directly applied to a membrane for detection. Its simplicity makes it applicable in various contexts, including monitoring labeling efficiency, estimating specific protein concentrations, and comparing antibody performance. For instance, to assess labeling efficiency with biotin, one would compare a labeled sample to a known biotinylated reference. This involves making serial dilutions of both the test sample and the reference standard, ideally starting with a 10-fold dilution to generate a broad range of samples.
