McClean: ConA - Revision history

Megan N McClean: /* Procedure */

Megan N McClean — Wed, 02 Nov 2011 22:04:19 GMT

Procedure

←Older revision		Revision as of 22:04, 2 November 2011
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	Mix 2.5mls of the CaCl<sub>2</sub> and the MnCl<sub>2</sub> solution in 45mls of water (50ml total volume).		Mix 2.5mls of the CaCl<sub>2</sub> and the MnCl<sub>2</sub> 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. 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.	+	In this mixture, dissolve a 20 mg/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 2 mg/ml solution to prime flow cells.

-	Final concentrations: 5mM MnCl<sub>2</sub> 5mM CaCl<sub>2</sub> ~~2mg~~/ml concanavalin A	+	Final concentrations: 5mM MnCl<sub>2</sub> 5mM CaCl<sub>2</sub> 2 mg/ml concanavalin A

	==Notes==		==Notes==

Megan N McClean: /* Procedure */

Megan N McClean — Wed, 02 Nov 2011 22:03:11 GMT

Procedure

←Older revision		Revision as of 22:03, 2 November 2011
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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.		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	+	Final concentrations: 5mM MnCl<sub>2</sub> 5mM CaCl<sub>2</sub> 2mg/ml concanavalin A

	==Notes==		==Notes==

Megan N McClean: /* Procedure */

Megan N McClean — Wed, 02 Nov 2011 22:02:26 GMT

Procedure

←Older revision		Revision as of 22:02, 2 November 2011
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	==Procedure==		==Procedure==
-	Mix 2.5mls of the ~~CaCl2~~ and the ~~MnCl2~~ solution in 45mls of water (50ml total volume).	+	Mix 2.5mls of the CaCl<sub>2</sub> and the MnCl<sub>2</sub> 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. 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.		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.

Megan N McClean: /* Materials */

Megan N McClean — Wed, 02 Nov 2011 22:00:48 GMT

Materials

←Older revision		Revision as of 22:00, 2 November 2011
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	* 1M CaCl<sub>2</sub> solution in sterile H<sub>2</sub>O		* 1M CaCl<sub>2</sub> solution in sterile H<sub>2</sub>O
	* 1M MnCl<sub>2</sub> solution in sterile H<sub>2</sub>O		* 1M MnCl<sub>2</sub> solution in sterile H<sub>2</sub>O
-	* Concanavalin A (MP Biomedicals, ~~cat no:~~ 150710)	+	* Concanavalin A (MP Biomedicals, #150710)

	==Procedure==		==Procedure==

Megan N McClean: /* Materials */

Megan N McClean — Wed, 02 Nov 2011 22:00:36 GMT

Materials

←Older revision		Revision as of 22:00, 2 November 2011
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	==Materials==		==Materials==
-	* 1M ~~CaCl2~~ solution in sterile ~~H20~~	+	* 1M CaCl<sub>2</sub> solution in sterile H<sub>2</sub>O
-	* 1M ~~MnCl2~~ solution in sterile ~~H20~~	+	* 1M MnCl<sub>2</sub> solution in sterile H<sub>2</sub>O
	* Concanavalin A (MP Biomedicals, cat no: 150710)		* Concanavalin A (MP Biomedicals, cat no: 150710)

Megan N McClean: /* Notes */

Megan N McClean — Wed, 02 Nov 2011 21:59:52 GMT

Notes

←Older revision		Revision as of 21:59, 2 November 2011
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	==Notes==		==Notes==
	Please feel free to post comments, questions, or improvements to this protocol. Happy to have your input! Please sign your name to your note by adding <font face="courier"><nowiki>'''*~~~~''':</nowiki></font> to the beginning of your tip.		Please feel free to post comments, questions, or improvements to this protocol. Happy to have your input! Please sign your name to your note by adding <font face="courier"><nowiki>'''*~~~~''':</nowiki></font> to the beginning of your tip.
-	*[[User:Megan N McClean\|Megan N McClean]] 17:59, 2 November 2011 (EDT) The solution can be stored at room temperature after the initial thaw for up to a week without noticeable loss of efficacy.	+	*[[User:Megan N McClean\|Megan N McClean]] 17:59, 2 November 2011 (EDT) The solution can be stored at room temperature after the initial thaw for up to a week without noticeable loss of efficacy. I have also put a solution of conA through several -20°C freeze thaws without noticing a significant change in "sticking power".

	==Contact==		==Contact==

Megan N McClean: /* Notes */

Megan N McClean — Wed, 02 Nov 2011 21:59:07 GMT

Notes

←Older revision		Revision as of 21:59, 2 November 2011
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	==Notes==		==Notes==
-	Please feel free to post comments, questions, or improvements to this protocol. Happy to have your input!	+	Please feel free to post comments, questions, or improvements to this protocol. Happy to have your input! Please sign your name to your note by adding <font face="courier"><nowiki>'''*~~~~''':</nowiki></font> to the beginning of your tip.
-	~~#List troubleshooting tips here.~~	+	*[[User:Megan N McClean\|Megan N McClean]] 17:59, 2 November 2011 (EDT) The solution can be stored at room temperature after the initial thaw for up to a week without noticeable loss of efficacy.
-	~~#You can also link to FAQs/tips provided by other sources such as the manufacturer or other websites.~~	+
-	~~#Anecdotal observations that might be of use to others can also be posted here.~~	+
-		+
-	Please sign your name to your note by adding <font face="courier"><nowiki>'''*~~~~''':</nowiki></font> to the beginning of your tip.	+

	==Contact==		==Contact==

Megan N McClean: /* Overview */

Megan N McClean — Tue, 11 Oct 2011 15:12:24 GMT

Overview

←Older revision		Revision as of 15:12, 11 October 2011
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	==Overview==		==Overview==
-	This is the concanavalin A solution we use for "sticking" yeast cells to the coverslip during microfluidics experiments.	+	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==		==Materials==

Megan N McClean: /* Contact */

Megan N McClean — Fri, 23 Sep 2011 21:12:50 GMT

Contact

←Older revision		Revision as of 21:12, 23 September 2011
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	==Contact==		==Contact==
-	*'''[[User:Megan N McClean\|Megan N McClean]] 14:01, ~~20 July~~ 2011 (EDT)'''	+	*'''[[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]].

Megan N McClean at 21:12, 23 September 2011

Megan N McClean — Fri, 23 Sep 2011 21:12:28 GMT

←Older revision		Revision as of 21:12, 23 September 2011
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	Final concentrations: 5mM MnCl2 5mM CaCl2 2mg/ml concanavalin A		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.
		+	#You can also link to FAQs/tips provided by other sources such as the manufacturer or other websites.
		+	#Anecdotal observations that might be of use to others can also be posted here.
		+
		+	Please sign your name to your note by adding <font face="courier"><nowiki>'''*~~~~''':</nowiki></font> to the beginning of your tip.
		+
		+	==Contact==
		+	*'''[[User:Megan N McClean\|Megan N McClean]] 14:01, 20 July 2011 (EDT)'''
		+
		+	or instead, [[Talk:{{PAGENAME}}\|discuss this protocol]].
		+
		+	<!-- You can tag this protocol with various categories. See the [[Categories]] page for more information. -->
		+
		+	<!-- Move the relevant categories above this line to tag your protocol with the label
		+	[[Category:Protocol]]
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