McClean: ConA

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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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Reagents:
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==Overview==
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• 1M CaCl2 solution in sterile H20
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This is the concanavalin A solution we use for "sticking" yeast cells to the coverslip during microfluidics experiments.
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• 1M MnCl2 solution in sterile H20
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• Concanavalin A (MP Biomedicals, cat no: 150710)
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==Materials==
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* 1M CaCl2 solution in sterile H20
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* 1M MnCl2 solution in sterile H20
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* Concanavalin A (MP Biomedicals, cat no: 150710)
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==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.
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In this mixture, dissolve a 20mg/ml solution of conA.
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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.
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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.
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Final concentrations: 5mM MnCl2 5mM CaCl2 2mg/ml concanavalin A

Revision as of 17:11, 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

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