4006184 Rev A

Bio-Rad Laboratories, 2000 Alfred Nobel Drive, Hercules CA 94547

Instruction Manual
Large Precast Gel
Product Information
Catalog
Number Product Description
161-1450 10% Tris-HCI, 193x183x1.0 mm, IPG comb*
161-1451 12% Tris-HCI, 193x183x1.0 mm, IPG comb*
161-1452 10–20% Tris-HCI, 193x183x1.0 mm, IPG comb*
161-1453 8–16% Tris-HCI, 193x183x1.0 mm, IPG comb
161-1454 10% Tris-HCI, 160x160x1.0 mm, 15 well comb
161-1455 12% Tris-HCI, 160x160x1.0 mm, 15 well comb
161-1456 10–20% Tris-HCI, 160x160x1.0 mm, 15 well comb
161-1457 8–16% Tris-HCI, 160x160x1.0 mm, 15 well comb
161-1458 10% Tris-HCI, 160x160x1.5 mm, 20 well comb
161-1459 12% Tris-HCI, 160x160x1.5 mm, 20 well comb
161-1460 10–20% Tris-HCI, 160x160x1.5 mm, 20 well comb
161-1461 8–16% Tris-HCI, 160x160x1.5 mm, 20 well comb
* Requires IPG Gaskets and Clamp Set
165-1835 IPG Clamp Set
165-3182 IPG Gaskets
1
 4006184 Rev A
Bio-Rad Laboratories, 2000 Alfred Nobel Drive, Hercules CA 94547

Instruction Manual
Large Precast Gel

Product Information
Catalog
Number Product Description
161-1450 10% Tris-HCI, 193x183x1.0 mm, IPG comb*
161-1451 12% Tris-HCI, 193x183x1.0 mm, IPG comb*
161-1452 10–20% Tris-HCI, 193x183x1.0 mm, IPG comb*
161-1453 8–16% Tris-HCI, 193x183x1.0 mm, IPG comb

161-1454 10% Tris-HCI, 160x160x1.0 mm, 15 well comb

161-1455 12% Tris-HCI, 160x160x1.0 mm, 15 well comb
161-1456 10–20% Tris-HCI, 160x160x1.0 mm, 15 well comb
161-1457 8–16% Tris-HCI, 160x160x1.0 mm, 15 well comb

161-1458 10% Tris-HCI, 160x160x1.5 mm, 20 well comb

161-1459 12% Tris-HCI, 160x160x1.5 mm, 20 well comb
161-1460 10–20% Tris-HCI, 160x160x1.5 mm, 20 well comb
161-1461 8–16% Tris-HCI, 160x160x1.5 mm, 20 well comb

* Requires IPG Gaskets and Clamp Set

165-1835 IPG Clamp Set
165-3182 IPG Gaskets

1
Introduction

For complete instructions, refer to the PROTEAN II Cells Instruction Manual.

1. Remove gel from package, being sure to hold the gel in a horizontal position, with the glass
plates parallel to the benchtop. Do not handle the gel in an upright, vertical position at this stage
as this may cause shifting of the gel between the plates. Do not set the bottom edges of the glass
plates on the benchtop.
2. Maintaining the horizontal position, attach the PROTEAN II sandwich clamps to either side of
the gel cassette so that the long plate fits snugly at the top of the clamps.
3. Remove the comb from the gel and rinse the wells with ddH2O or electrode buffer.
4. With the cooling core flat on the benchtop, wet the U-shaped gasket with ddH2O or upper
electrode buffer solution to ensure a good seal.
5. With the short glass plate facing the cooling core, position the sandwich so that the grooves in
the upper portion of the clamps are fitted onto the locating pins on the central cooling core. The
gel sandwich should be positioned at an angle of ≤ 20° with the cooling core.
6. With your fingers below the latch on the cooling core and your thumbs resting on the clamps,
gently push the gel sandwich onto the cooling core in one fluid motion. The upper edge of the
short inner glass plate should be butted against the notches of the U-shaped gasket, and the tabs
of each clamp should be held securely against the latch assemblies on either side of the cooling
core.
7. Turn the cooling core to its other side and repeat all steps to attach the second gel sandwich.
8. Pour 350 ml of electrode buffer into the upper buffer chamber. Check the integrity of the upper
buffer seal.
9. Pour enough electrode buffer into the lower buffer chamber to cover at least the bottom 1 cm
of the gel (approximately 1200 ml for 20 cm gels, and 1800 ml for 16 cm gels). Lower the
central cooling core into the lower buffer chamber at a slight angle to prevent trapping air
bubbles under the gel. A few, isolated bubbles will not affect the run.
10. Load samples.
11. Place lid on top of the assembly and attach the leads to a suitable power supply. Run gel(s) at
constant current. Refer to the chart below for running conditions.
Gel Thickness Stacking Gel Run Time Separating Gel Run Time

1.0 mm 16 mA/gel 30 min 24 mA/gel ~ 4 hrs, 40 min

1.5 mm 25 mA/gel 30 min 35 mA/gel ~ 2 hrs, 30 min