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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 without transferring samples from a gel. Instead, samples are dotted directly onto a membrane and probed for detection. Its simplicity allows for various applications, including monitoring labeling efficiency, estimating protein concentration, and comparing antibody performance. For example, to assess labeling efficiency with biotin, one would compare the labeled sample to a known biotinylated reference standard. This involves creating serial dilutions of both the test sample and reference standard in the same manner, with a 10-fold dilution recommended to cover a broad range of samples.
