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Dot blot is a straightforward method for detecting biological samples, including proteins and nucleic acids. It operates on principles similar to western and southern blotting, but instead of transferring samples from a gel, they are directly dotted onto a membrane for detection. Due to its simplicity, dot blot is versatile and used in various applications, such as monitoring labeling efficiency, estimating protein concentration, and comparing antibody performance. For example, determining labeling efficiency with a biotin-labeled sample involves comparing it to a known biotinylated reference standard. This can be done by creating serial dilutions of both the test sample and the reference, ensuring identical dilution processes for accurate comparisons.
