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Dot blot is a straightforward method for detecting biological samples, such as proteins or nucleic acids, similar to western and southern blotting, but samples are applied directly onto a membrane instead of being blotted from a gel. Its simplicity allows for various applications, including monitoring labeling efficiency with different probes, estimating protein concentration, and comparing antibody performance. For instance, to determine labeling efficiency with a biotin-labeled sample, one should compare it to a known biotinylated reference standard by creating serial dilutions of both the test sample and the reference, ensuring identical dilution methods, with a 10-fold serial dilution being recommended for a broad range of samples.
