SDS-PAGE
From OpenWetWare
(Difference between revisions)
(→Solutions for 10 ml (MINIgel) of various polyacrylamide gels (SDS-PAGE)) |
Current revision (10:48, 8 April 2009) (view source) (SDS link) |
||
| (6 intermediate revisions not shown.) | |||
| Line 1: | Line 1: | ||
| - | === | + | ===Running (lower) gel: 10 ml (2 MINIgels)=== |
| + | |||
Chemical 8% 10% 12% | Chemical 8% 10% 12% | ||
| Line 5: | Line 6: | ||
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]] | ||
Current revision
Contents |
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.
