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Dot blot is a straightforward technique for detecting biological samples, such as proteins or nucleic acids. It operates on principles similar to western and southern blotting but involves directly dotting samples onto a membrane for detection, rather than transferring from a gel. Due to its simplicity, dot blot is versatile and supports various applications, including monitoring labeling efficiency, estimating protein concentration, and comparing antibody performance. For instance, to assess labeling efficiency of a biotin-labeled sample, one would create serial dilutions of both the test sample and a known biotinylated reference standard, ensuring both are diluted identically, often starting with a 10-fold serial dilution for a broad concentration range.
