Bio-Rad ReadyPrep™ 2-D Starter Kit User 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