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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 09:24, 14 June 2022
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=== Solutions === | === Solutions === | ||
====Stain==== | ====Stain==== | ||
<b>0.2-0.3 % Coomassie R-250, 10 % methanol, 10 % acetic acid, one liter:</b> | Old: <b>0.2-0.3 % Coomassie R-250, 10 % methanol, 10 % acetic acid, one liter:</b> | ||
Weigh about 2-3 grams of Coomassie and place into a glass bottle. | Weigh about 2-3 grams of Coomassie and place into a glass bottle. | ||
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Carefully add glacial acetic acid to make the liter volume. The acid promotes Coomassie aggregation and sticking to other surfaces (and proteins!), so add it last to help the Coomassie dissolve well. | Carefully add glacial acetic acid to make the liter volume. The acid promotes Coomassie aggregation and sticking to other surfaces (and proteins!), so add it last to help the Coomassie dissolve well. | ||
New: The above is a common concentration of Coomassie, but it's really way too much and causes more debris to coat the gels. It can be prepared at 1/5 to 1/10 that (0.04 - 0.02 %) and it works fine. | |||
====Destain==== | ====Destain==== | ||
