Analysis of Protein Posttranslational Modifications Using DIGE-Based Proteomics - Difference Gel Electrophoresis (DIGE): Methods and Protocols - page 139

4. Wash strips in DTT-EB. Use 10 mL of DTT-EB for 7-cm strips
and 20 mL for 13- and 24-cm strips. Wrap tubes in aluminum
foil and place them on an orbital shaker for 10 min.
5. Rinse strips in 1× running buffer, then transfer to fresh tubes
with Iodo-EB. Again, use 10 mL for 7-cm strips and 20 mL for
13- and 24-cm strips. Wrap tubes in aluminum foil and place
them on an orbital shaker for 10 min.
6. Rinse each strip in 1× running buffer and then load each strip
onto a gel. Lay the top of the gel down onto a rack for 1.5-mL
tubes with the long plate on the bottom. Place the nongel side
of the strip on the long plate side of the gel that is facing up
(Fig. 3 ). Carefully push the strip down between the glass plates,
Fig. 3. Loading the strip onto the gel. ( a ) Lay the strip down on the long glass plate with the gel side facing up. ( b ) Using
fi ne gel loading tips, push the strip down between the glass plates until it meets the acrylamide gel.