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Dot blot is a straightforward method for detecting biological samples like proteins or nucleic acids. It operates on principles similar to western and southern blotting but differs in that samples are directly dotted onto a membrane rather than blotted from a gel. Due to its simplicity, dot blot supports various applications, including monitoring labeling efficiency, estimating protein concentration, and comparing antibody performance. For instance, to determine labeling efficiency of a biotin-labeled sample, one would compare it to a known biotinylated reference standard. This involves making serial dilutions of both the test sample and reference standard, using a consistent dilution method, with a 10-fold serial dilution being recommended for a wide range of samples.
