SDS-PAGE

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

Resources

SDS-PAGE "Hall of Shame" - examples of common failures with debugging tips.

• Lidstrom: SDS-PAGE
• protein gel electrophoresis tips and troubleshooting guide (download)] [[Category:PAGE]