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Dot blot is a straightforward method for detecting biological samples like proteins or nucleic acids, resembling western and southern blotting but without the transfer from a gel. Instead, samples are directly dotted onto a membrane for probing. Its simplicity allows for various applications, including monitoring labeling efficiency, estimating protein concentration, and comparing antibody performance. For instance, to assess labeling efficiency, a sample labeled with biotin is compared to a known biotinylated reference standard. This involves creating serial dilutions of both the test sample and reference standard, ensuring identical dilution processes, with a 10-fold dilution serving as a practical initial approach for a broad range of samples.
