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After Western blot transfer, the membrane is prepared for protein probing through a series of incubation and wash steps. Blocking is the first step, where a non-reactive protein coats the membrane to prevent non-specific antibody binding. Nitrocellulose and PVDF membranes are ideal for high protein-binding affinity. This prevents high background signals that can hinder protein detection. The blocking buffer typically contains non-reactive proteins like BSA, non-fat milk, or serum, and Tris- or phosphate-buffered saline with Tween-20 detergent.
