This is the concanavalin A solution we use for "sticking" yeast cells to the coverslip during microfluidics experiments.
- 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 -20°C, thaw just before microscopy. Use the 2mg/ml solution to prime flow cells.
Final concentrations: 5mM MnCl2 5mM CaCl2 2mg/ml concanavalin A
Please feel free to post comments, questions, or improvements to this protocol. Happy to have your input!
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- Megan N McClean 14:01, 23 September 2011 (EDT)
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