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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 for probing and detection. Due to its simplicity, dot blot is used in various applications, including monitoring labeling efficiency, estimating specific protein concentrations, and comparing different antibodies. For instance, to assess labeling efficiency with biotin, one compares a labeled sample to a known biotinylated reference standard. This is done by preparing serial dilutions of both the test sample and reference standard, ensuring identical dilutions, starting with a 10-fold series for broad range assessment.
