Bio-Rad PROTEAN® i12™ IEF System - Manual (Page 14)

PROTEAN i12 IEF System Instruction Manual 10 |
Chapter 2 Basic Operation
Fig. 2.3. Placement of an IPG strip gel-side up into the i12 focusing tray.
3. Overlay each IPG strip with mineral oil to prevent evaporation and precipitation of urea during rehydration
(see Table 2.1 for recommended volumes). Apply the mineral oil to both ends of the channel and allow it
to flow toward the middle of the channel. IPG strips can be left to rehydrate for up to 1 hr before adding
the mineral oil.
4. Cover the tray and leave it on a level bench overnight (12–18 hr) for complete rehydration.
5. Transfer the rehydrated IPG strips to the focusing tray for IEF (see below).
IEF with Gel-Side Up
For IEF of IPG strips that were rehydrated in the presence of sample (in-gel loading):
1. Using forceps, remove the IPG strips from the rehydration tray, remove excess mineral oil, and place the
rehydrated IPG strips gel-side up in the channels of the focusing tray (Figure 2.3, A). Position the positive
(+) ends of the IPG strips against the positioning stops in each channel.
2. (Recommended) Wet the gel-side up wicks (notched) with distilled or deionized water and blot off excess
water. Use two wicks per IPG strip: place a wick at each end of each IPG strip (Figure 2.3, B).
3. Position the electrode assemblies in the focusing tray and press down on the green tabs to snap the
electrode assemblies into place (Figure 2.3, C). Place the focusing tray with the rehydrated IPG strips on
the Peltier platform and connect the electrodes to the instrument (see Section 2.1, Setup).
4. Overlay each IPG strip with mineral oil (see Table 2.1 for recommended volumes).
5. Select or program the protocol(s) and start the run (see Section 2.3, Starting the Run).
Place IPG strips gel-side up in channel of focusing tray. A
(Recommended) Place wet electrode
wicks on both ends of the IPG strip. B
Place electrode assemblies in
the focusing tray. Press down
on the green tabs to snap the
assemblies into place. C
IPG strip positioning stop