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Dot blot is a straightforward method for detecting biological samples, such as proteins or nucleic acids. It is similar to western and southern blotting, but instead of transferring samples from a gel, samples are directly dotted onto a membrane for detection. Due to its simplicity, dot blot is employed in various applications, including monitoring labeling efficiency, estimating protein concentrations, and comparing antibody performance. For instance, to assess labeling efficiency with 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, typically using a 10-fold dilution to achieve a broad range of concentrations.
