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Dot blot is a straightforward method for detecting biological samples like proteins or nucleic acids, functioning similarly to western and southern blotting but without the need for gel transfer. Instead, samples are dotted directly onto a membrane for probing. This technique is versatile and supports various applications, including monitoring labeling efficiency, estimating protein concentrations, and comparing antibody performance. For example, to determine labeling efficiency with biotin-labeled samples, one would compare the labeled sample to a known biotinylated reference standard. This involves creating serial dilutions of both the test sample and the reference, ensuring the same dilution method is used, with a 10-fold dilution being a common starting point for a broad range of samples.