SDS-PAGE: Difference between revisions
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=== | ===Running (lower) gel: 10 ml (2 MINIgels)=== | ||
Chemical 8% 10% 12% | Chemical 8% 10% 12% | ||
Water 4.74 4.0 3.3 ml | Water 4.74 4.0 3.3 ml | ||
30% AA(*) 2.66 3.3 4.0 ml | 30% AA(*) 2.66 3.3 4.0 ml | ||
LOWER buffer 2.5 2.5 2.5 ml | LOWER buffer 2.5 2.5 2.5 ml | ||
20% [[SDS]] 50 50 50 ul | |||
10% APS 200 200 200 ul | |||
TEMED 8.2 8.2 8.2 ul | |||
===Stacking (upper) gel: 4 ml (2 MINIgels)=== | |||
Chemical 4% | |||
Water 2.65 ml | |||
30% AA(*) 0.67 ml | |||
UPPER buffer 0.65 ml | |||
20% [[SDS]] 20 ul | |||
10% APS 125 ul | |||
TEMED 6 ul | |||
===Buffers=== | |||
LOWER buffer: 1M Tris base, pH 8.8 | |||
UPPER buffer: 1M Tris base, pH 6.8 | |||
===Abbreviations=== | |||
AA = acrylamide | |||
[[SDS]] = sodium dodecylsulfate | |||
APS = ammonium persulfate | |||
==Notes== | |||
[http://www.ruf.rice.edu/~bioslabs/studies/sds-page/sdsgoofs.html SDS-PAGE "Hall of Shame"] - examples of common failures with debugging tips. | |||
[[Category:Material]] [[Category:PAGE]] | |||
Revision as of 07:48, 8 April 2009
Running (lower) gel: 10 ml (2 MINIgels)
Chemical 8% 10% 12% Water 4.74 4.0 3.3 ml 30% AA(*) 2.66 3.3 4.0 ml LOWER buffer 2.5 2.5 2.5 ml 20% SDS 50 50 50 ul 10% APS 200 200 200 ul TEMED 8.2 8.2 8.2 ul
Stacking (upper) gel: 4 ml (2 MINIgels)
Chemical 4% Water 2.65 ml 30% AA(*) 0.67 ml UPPER buffer 0.65 ml 20% SDS 20 ul 10% APS 125 ul TEMED 6 ul
Buffers
LOWER buffer: 1M Tris base, pH 8.8 UPPER buffer: 1M Tris base, pH 6.8
Abbreviations
AA = acrylamide SDS = sodium dodecylsulfate APS = ammonium persulfate
Notes
SDS-PAGE "Hall of Shame" - examples of common failures with debugging tips.
