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Dot blot is a method for detecting biological samples. It is similar to Western and Southern blotting but samples are directly dotted onto a membrane. It is used for monitoring labeling efficiency, estimating protein concentration, and comparing antibody performance. To determine labeling efficiency with biotin, compare the labeled sample to a known biotinylated reference standard by making serial dilutions of both samples. A 10-fold dilution is a good starting point for a wide range of samples.