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Sauer:bis-Tris SDS-PAGE, the very best: Difference between revisions
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Sauer:bis-Tris SDS-PAGE, the very best (view source)
Revision as of 08:41, 31 January 2020
, 31 January 2020→Procedure
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Drain away the water (without dropping your gel in the sink), and cover with ~0.5 cm of strain solution. | Drain away the water (without dropping your gel in the sink), and cover with ~0.5 cm of strain solution. | ||
Place the gel in stain in the microwave and microwave on high until the solution just begins to boil (this step greatly accelerates the procedure and allows you to see you | Place the gel in stain in the microwave and microwave on high until the solution just begins to boil (this step greatly accelerates the procedure and allows you to see you bands in a minute or so). | ||
Remove from the microwave and gently shake or rock for a few minutes. Once you see the gel filled with Coomassie, it's done. | Remove from the microwave and gently shake or rock for a few minutes. Once you see the gel filled with Coomassie, it's done. | ||
Drain the Coomassie away and cover with Destain solution and a couple of Kim-wipes folded flat over the gel. | Drain the Coomassie away and cover with water, rock for about 5 minutes, drain. | ||
Cover with Destain solution and a couple of Kim-wipes folded flat over the gel. | |||
Microwave again on high unit l the solution begins to boil. | Microwave again on high unit l the solution begins to boil. | ||
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The gel should be clear with dark purple protein bands. | The gel should be clear with dark purple protein bands. | ||
If there is a Coomassie film on the surface, first image your gel, then try wiping the film off with a clean Kim-wipe before getting another image. This step can cause your gel to | If there is a Coomassie film on the surface, first image your gel, then try wiping the film off with a clean Kim-wipe before getting another image. This step can cause your gel to split, so it's better to get the image first. | ||
