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Dot blot is a straightforward method for detecting proteins or nucleic acids, similar to western and southern blotting, but with samples directly dotted onto a membrane for probing. Due to its simplicity, dot blot is versatile and supports various applications, including monitoring labeling efficiency, estimating protein concentration, and comparing antibody performance. For example, to determine labeling efficiency with biotin, one would compare a labeled sample to a known biotinylated reference standard. This involves making serial dilutions of both the test sample and the reference, ensuring they are diluted in the same manner, with a 10-fold serial dilution recommended for a broad range of samples.
