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Dot blot is a quick and straightforward method for detecting biological samples like proteins or nucleic acids. It shares principles with western and southern blotting but differs as samples are directly dotted onto a membrane instead of blotted from a gel. This simplicity allows it to support various applications, including monitoring labeling efficiency, estimating protein concentration, and comparing different antibodies. For instance, to determine labeling efficiency with biotin, one compares a labeled sample to a known biotinylated reference standard by creating serial dilutions of both the test sample and reference, ensuring identical dilution methods, with a 10-fold serial dilution providing a suitable range of samples.
