Bio-Rad ReadyPrep™ 2-D Starter Kit - Manual (Page 12)

Draining the oil essentially washes the
outside surface of the gel, removing
unabsorbed protein and resulting in
reduced horizontal streaking. If desired,
the disposable rehydration/equilibration
tray can be cleaned, dried, and used to
store strips after the completion of the
first-dimension run.
5 Cover each IPG strip with 2 to 3 ml of
fresh mineral oil. Check for, and if appli-
cable remove, any trapped air bubbles
beneath the strips. Place the lid onto the
tray (positive “+” to the left when the
inclined portion of the tray is on the right
as in Figure 8).
6 Place the focusing tray into the PROTEAN IEF cell and close the cover.
7 Program the PROTEAN IEF cell using the appropriate 3-step protocol in Table 2. For all
strip lengths, use the default cell temperature of 20ºC, with a maximum current of 50
µA/strip and No Rehydration. Refer to Section 5, Appendix, or the PROTEAN IEF cell
manual for details on entering the run parameters.
8 Press START to initiate the electrophoresis run.
10
Remember to observe the correct polarity
during the transfer of strips into the focus-
ing tray. The convenient “+” and “pH4-7”
printed on the strips (acidic end) should be
positioned at the end of the tray marked
with the “+” (this is the left side of the tray
when the inclined portion of the tray is on
the right as in Figure 7)
7 cm Voltage Time Volt-Hours Ramp
Step 1 250 20 min ----- Linear
Step 2 4,000 2 hr ----- Linear
Step 3 4,000 ----- 10,000 V-hr Rapid
Total 5 hr 14,000 V-hr
Table 2.
11 cm Voltage Time Volt-Hours Ramp
Step 1 250 20 min ----- Linear
Step 2 8,000 2.5 hr ----- Linear
Step 3 8,000 ----- 20,000 V-hr Rapid
Total 5.3 hr ~30,000 V-hr
17 cm Voltage Time Volt-Hours Ramp
Step 1 250 20 min ----- Linear
Step 2 10,000 2.5 hr ----- Linear
Step 3 10,000 ----- 40,000 V-hr Rapid
Total 7 hr ~50,000 V-hr