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Dot blot is a simple method for detecting biological samples like proteins or nucleic acids, akin to western and southern blotting, but involves directly dotting samples onto a membrane rather than blotting from a gel. Its simplicity allows for various applications, such as monitoring labeling efficiency, estimating protein concentration, and comparing antibody performance. For example, to determine labeling efficiency of a biotin-labeled sample, one should compare it with a known biotinylated reference standard. This involves creating serial dilutions of both the test sample and the reference standard in the same manner, with a 10-fold serial dilution recommended to provide a broad range of samples for analysis.
