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Dot blot is a straightforward method for detecting biological samples like proteins or nucleic acids. It operates similarly to western and southern blotting, but instead of transferring samples from a gel, they are directly spotted onto a membrane for detection. Due to its simplicity, dot blot is applicable in various scenarios, such as monitoring labeling efficiency, estimating protein concentration, and comparing antibody performance. For instance, to assess labeling efficiency in a biotin-labeled sample, one would compare it to a known biotinylated reference standard. This involves creating serial dilutions of both the test sample and reference standard, ideally starting with a 10-fold dilution for a broad range.
