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20. A step 5 of 200 V for 24 h may be added as a “hold” step.
21. If step 5 is applied for more than 6 h, apply a further step of
8,000 V for 1 h before proceeding.
22. If bubbles are introduced between the strip and acrylamide
gel, tapping the back of the strip (plastic) is usually enough to
remove them. If not, lift the strip from one end and begin
again, gently pushing the strip down at an angle so that bub-
bles are squeezed out toward the free end.
Acknowledgments
This work was supported by the UNC Systems-Proteomics Center
and the Duke Neuroproteomics Center.
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