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Dot blot is a simple method for detecting biological samples such as proteins or nucleic acids. It operates on principles similar to western and southern blotting, but samples are dotted directly onto a membrane for probing, without prior gel blotting. Due to its simplicity, dot blot is widely used in various applications, including monitoring labeling efficiency, estimating protein concentration, and comparing antibody performance. For example, to determine labeling efficiency with biotin, a user compares a test sample with a known biotinylated reference by making serial dilutions of both, ensuring identical dilution processes. A 10-fold serial dilution is recommended for a sufficient range of samples.
