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Dot blot is a straightforward method for detecting biological samples like proteins or nucleic acids, following principles similar to western and southern blotting. Unlike those methods, samples are directly dotted onto a membrane for detection rather than being blotted from a gel. Its simplicity makes it applicable for various tasks, such as monitoring labeling efficiency with different probes, estimating specific protein concentrations, and comparing antibody performance. For instance, to determine labeling efficiency of a biotin-labeled sample, one compares it to a known biotinylated reference standard by making serial dilutions of both the test sample and reference, ensuring equal dilution methods, with a 10-fold serial dilution being effective for a wide range of samples.
