McClean: ConA: Difference between revisions
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(New page: 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 4...) |
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==Overview== | |||
This is the concanavalin A solution we use for "sticking" yeast cells to the coverslip during microfluidics experiments. | |||
==Materials== | |||
* 1M CaCl2 solution in sterile H20 | |||
* 1M MnCl2 solution in sterile H20 | |||
* Concanavalin A (MP Biomedicals, cat no: 150710) | |||
==Procedure== | |||
Mix 2.5mls of the CaCl2 and the MnCl2 solution in 45mls of water (50ml total volume). | 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. | 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. | 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. | ||
Dilute from this mixture down to 2mg/ml in miliQ water. Store the 2mg/ml aliquots at - | |||
Final concentrations: 5mM MnCl2 5mM CaCl2 2mg/ml concanavalin A | |||
Revision as of 14:11, 23 September 2011
Overview
This is the concanavalin A solution we use for "sticking" yeast cells to the coverslip during microfluidics experiments.
Materials
- 1M CaCl2 solution in sterile H20
- 1M MnCl2 solution in sterile H20
- Concanavalin A (MP Biomedicals, cat no: 150710)
Procedure
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
