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Dot blot is a straightforward method for detecting biological samples such as proteins or nucleic acids. It operates similarly to western and southern blotting, but instead of transferring samples from a gel, the samples are directly dotted onto a membrane for detection. Due to its simplicity, dot blot is applied in various contexts, including monitoring labeling efficiency, estimating protein concentration, 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 making serial dilutions of both the sample and the reference standard, with a 10-fold serial dilution offering a broad range of samples for analysis.
