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Dot blot is a straightforward method for detecting biological samples such as proteins or nucleic acids, similar in principle to western blotting and southern blotting, but instead of transferring samples from a gel, they are directly dotted onto a membrane for detection. Its simplicity makes it suitable for various applications, including monitoring labeling efficiency with different probes, estimating protein concentration, and comparing antibody performance. For instance, to assess labeling efficiency of a biotin-labeled sample, serial dilutions of the test sample and a known biotinylated reference standard are prepared, ensuring identical dilution methods, with a 10-fold serial dilution being optimal for obtaining a comprehensive range of samples.
