A protein-free blocking system for the covalently binding matrix cyanuric chloride-activated paper in immunological...

A new protein-free blocking system containing 10% (w/v) sulfanilic acid/10% (v/v) triethanolamine/0.05% (v/v) Tween 20 has been used to block free binding sites of the covalently binding matrix cyanuric chloride-activated paper (CCA-paper). This method allows a reversible staining of protein blots with Coomassie brilliant blue after each step of the immunological procedure and reuse of the blots for a repeated antibody probing. Non-radioactive and radioactive detection procedures have been compared with blots on CCA-paper and nitrocellulose. The best combination is a Coomassie brilliant blue staining and immunological detection with 125I-protein A.