McClean: ConA

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Revision as of 10:12, 11 October 2011 by Megan N McClean (Talk | contribs)
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Contents

Overview

This is the concanavalin A solution we use for "sticking" yeast cells to the coverslip during microfluidics experiments. Generally, we make a large stock and store it in the -20°C freezer in ~ 500μL aliquots. These can be removed from the freezer and thawed at room temperature when needed. For most applications, the thawed stock can be left at room temperature for several days (or even freeze-thawed several times) before it's efficiency is reduced noticeably.

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

Notes

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