Knight:NuPAGE electrophoresis/Hybrid staining - Revision history

Reshma P. Shetty: /* Procedure */

Reshma P. Shetty — Thu, 23 Aug 2007 16:25:13 GMT

Procedure

←Older revision		Revision as of 16:25, 23 August 2007
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	#Gel can be stored in salt solution for several weeks.		#Gel can be stored in salt solution for several weeks.

-	[[Category:Protein]] [[Category:In vitro]]	+	[[Category:Protein]] [[Category:In vitro]] [[Category:Protocol]]

Reshma P. Shetty at 20:15, 1 June 2007

Reshma P. Shetty — Fri, 01 Jun 2007 20:15:21 GMT

←Older revision		Revision as of 20:15, 1 June 2007
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	#*''Only do this step if you aren't planning on drying the gel! Otherwise, when the gel dries, you will see a white salt precipitate.''		#*''Only do this step if you aren't planning on drying the gel! Otherwise, when the gel dries, you will see a white salt precipitate.''
	#Gel can be stored in salt solution for several weeks.		#Gel can be stored in salt solution for several weeks.
		+
		+	[[Category:Protein]] [[Category:In vitro]]

Reshma P. Shetty at 23:43, 16 January 2007

Reshma P. Shetty — Tue, 16 Jan 2007 23:43:23 GMT

New page

==Overview==
A fast protocol for visualizing bands on a polyacrylamide gel via coomassie-like staining.

==Materials==
*Deionized water
*[http://www.invitrogen.com/content.cfm?pageid=9869 Invitrogen SimplyBlue SafeStain]
*Microwave
*Plastic tray
**''Use lid of a 1000μL pipette tip box.''
*Orbital shaker
*Kimwipes

==Procedure==
#Add 100mL deionized water in staining tray.
#Microwave staining tray loosely covered for 30 secs on high.
#Shake staining tray for 1 min on orbital shaker at room temperature.
#Discard water.
#Repeat steps 1-4 two more times.
#Add 20mL SimplyBlue SafeStain (enough to just cover the gel).
#Microwave staining tray loosely covered for 20 secs on high.
#Shake staining tray for 5 mins on orbital shaker at room temperature.
#*''You should now be able to see most of the bands on the gel.''
#Place staining tray with gel inside a ziploc bag to limit evaporation.
#Incubate overnight on orbital shaker at room temperature.
#*''Overnight incubation will yield better sensitivity due to darker staining.''
#Discard SimplyBlue SafeStain.
#Add 100mL deionized water.
#Add 2 kimwipes.
#Shake staining tray for 2-3 hours on orbital shaker at room temperature.
#Take gel picture.
#Add 20mL 20% NaCl for at least 5 mins.
#*''Only do this step if you aren't planning on drying the gel! Otherwise, when the gel dries, you will see a white salt precipitate.''
#Gel can be stored in salt solution for several weeks.