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Dot blot is a simple method for detecting biological samples like proteins or nucleic acids, similar to western and southern blotting but without the need for gel transfer. Instead, samples are directly dotted 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 determine labeling efficiency of a biotin-labeled sample, one would compare it to a known biotinylated reference standard. This involves making serial dilutions of both the test sample and the reference, ensuring they are diluted equivalently, with a 10-fold serial dilution being effective for producing a wide range of samples.
