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Dot blot is a straightforward method for detecting biological samples such as proteins or nucleic acids. It operates similarly to western and southern blotting, but instead of transferring samples from a gel, they are dotted directly onto a membrane for probing. Due to its simplicity, dot blot is versatile, supporting various applications including monitoring labeling efficiency, estimating protein concentration, and comparing antibody performance. For example, to determine labeling efficiency with biotin, the labeled sample is compared to a known biotinylated standard. This involves creating serial dilutions of both the test sample and the reference standard, with a 10-fold serial dilution providing a broad range for analysis.
