The gels were
run at 120V during 10 min and after that at 180V until the advancing
front reached the button of the gel. For 2D-PAGE, 300 g of
protein were resuspended in 50 L of lysis buffer (9.5 M urea, 2%
Triton X-100, 1.6% ampholytes 4–7 range, 0.4% ampholytes 3–10
range, and 5% -mercaptoethanol), incubated at room temperature
for 15 min and loaded onto lab-made first dimension gels (115 mm
height and 3 mm internal diameter capillary tubes). A 4