Bio-Rad PROTEAN® IEF Cell Accessories - Manual (Page 7)

2.2 IPG Strip Rehydration Solution Preparation and Strip Rehydration
1. Prepare rehydration sample buffer and rehydrate IPG strips just prior to sample cup-loading
and isoelectric focusing. Add the indicated volume of rehydration buffer to each of the
required channels in the rehydration tray to rehydrate to 0.53 mm thickness.
Note: Rehydration to 0.53 mm thickness ensures a complete seal between the cup and the
IPG strip. The following volumes are 10% greater than those recommended in the ReadyStrip
IPG Strip instruction manual.
IPG Length 7 cm 11 cm 17 cm 18 cm 24 cm
Rehydration
volume for
135 µl 200 µl 330 µl 345 µl 450 µl
cuploading
2. Carefully remove the IPG strip cover sheet and place the IPG strip gel side down in the
rehydration trays. To ensure even rehydration of the IPG strips, the entire surface to the IPG
strips must be completely wetted.
Figure 4. Removal of IPG strip cover sheet.
3. Overlay IPG strips with mineral oil and rehydrate strips ~ 12 hours (or overnight). To maintain a
constant temperature during rehydration, place the rehydration tray on the cooling platform of
the PROTEAN IEF cell and set-up the rehydration program. (See PROTEAN IEF cell manual
for details).
2.3 Transfer and Placement of Rehydrated IPG Strips in the Focusing Tray
1. Remove the rehydrated IPG strips from the rehydration tray. Remove excess mineral oil by
carefully blotting the IPG strip on filter paper wetted with DI water.
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