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Dot blot is a simple method for detecting biological samples such as proteins or nucleic acids, akin to western and southern blotting, but involves directly dotting samples onto a membrane instead of transferring from a gel. Its simplicity allows for various applications, 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 test sample to a known biotinylated reference standard. This process involves making serial dilutions of both the test sample and the reference standard in the same manner, starting with a 10-fold dilution to create a broad concentration range.
