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Dot blot is an efficient method for detecting biological samples like proteins or nucleic acids, functioning similarly to western and southern blotting. However, in dot blotting, samples are directly applied to a membrane instead of being transferred from a gel, allowing for easier detection. This simplicity makes dot blot versatile for various applications, including monitoring labeling efficiency with different probes, estimating specific protein concentrations, and comparing antibody performance. For instance, to assess labeling efficiency of a biotin-labeled sample, one would compare it with a known biotinylated reference standard by performing serial dilutions on both, starting with a 10-fold dilution to achieve a broad range of samples.