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The ideal blocking buffer depends on your protein and detection method. BSA works well for phosphorylated proteins or biotin-avidin systems, while milk may interfere with some assays.
Phosphate-Buffered Saline (PBS) is isotonic and non-toxic, mimicking physiological ion concentrations, making it suitable for washing and antibody incubation in immunoassays.
PBS and TBS are often interchangeable. Use TBS when detecting phosphorylated proteins to avoid phosphate interference with phospho-specific antibodies.
Low concentrations of Tween-20 (0.05–0.1%) in PBS or TBS help remove unbound antibodies, reducing background and improving signal clarity.