McClean: ConA: Difference between revisions
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==Contact== | ==Contact== | ||
*'''[[User:Megan N McClean|Megan N McClean]] 14:01, | *'''[[User:Megan N McClean|Megan N McClean]] 14:01, 23 September 2011 (EDT)''' | ||
or instead, [[Talk:{{PAGENAME}}|discuss this protocol]]. | or instead, [[Talk:{{PAGENAME}}|discuss this protocol]]. | ||
Revision as of 14:12, 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
Notes
Please feel free to post comments, questions, or improvements to this protocol. Happy to have your input!
- List troubleshooting tips here.
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- Anecdotal observations that might be of use to others can also be posted here.
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Contact
- Megan N McClean 14:01, 23 September 2011 (EDT)
or instead, discuss this protocol.
