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Dot blot is a straightforward method for detecting biological samples, such as proteins or nucleic acids. It resembles western and southern blotting but involves applying samples directly onto a membrane without prior gel transfer. Due to its simplicity, dot blot is versatile for various applications, including monitoring labeling efficiency, estimating protein concentration, and comparing antibody performance. For assessing labeling efficiency with biotin, one compares a labeled sample to a known biotinylated reference standard. This process entails making serial dilutions of both the test sample and reference standard, ensuring identical dilution methods, with a 10-fold serial dilution providing a broad range for analysis.
