Reagents: • 1M CaCl2 solution in sterile H20 • 1M MnCl2 solution in sterile H20 • Concanavalin A (MP Biomedicals, cat no: 150710)
Mix 2.5mls of the CaCl2 and the MnCl2 solution in 45mls of water (50ml total volume). Check the pH of this solution. It should be between 6-7. Do not pH this solution with NaOH as the manganese will precipitate out. In this mixture, dissolve a 20mg/ml solution of conA. Dilute from this mixture down to 2mg/ml in miliQ water. Store the 2mg/ml aliquots at -20C, thaw just before microscopy. Use the 2mg/ml solution to prime flow cells.