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Semi-Wet Blotting of E-PAGE™ Gels
Introduction
Semi-Wet
Transfer
Apparatus
Important
A semi-wet blotting procedure using an XCell II™ Blot Module for transfer of
proteins from E-PAGE™ Gels is described below.
Two XCell II™ Blot Module units are required to transfer one E-PAGE™ Gel.
To obtain the best transfer, we recommend transferring only one half of an
E-PAGE™ Gel in one XCell II™ Blot Module. Inefficient transfer may occur if you
transfer 2 halves of the E-PAGE™ Gel in one blot module at the same time, as the
E-PAGE™ Gel is thicker than regular mini-gels.
If you are using the XCell II™ Blot Module for transferring one E-PAGE™ Gel, we
recommend that you have two XCell II™ Blot Modules on hand before
proceeding for transfer.
Other semi-wet transfer apparatus that can accommodate an E-PAGE™ Gel
(8.6 cm x 13.5 cm) may be used. However you may get sub-optimal results if
your blotting apparatus is prone to transfer distortion when accommodating
thick gels. It may be necessary to optimize your semi-wet blotting procedure
based on the protocol below, or it may be necessary to use a semi-dry blotting
apparatus to obtain optimal results.
Note: Using the Criterion Blotter module from BioRad (BioRad Cat. no.
170-4070) is not recommended.
Materials Needed
You will need the following items. Ordering information is on page 71.

XCell II™ Blot Module (2 units required for transfer of 1 E-PAGE™ Gel)

XCell SureLock™ Mini-Cell

Power supply (1–2 amps)

Methanol

NuPAGE® Transfer Buffer (20X)

NuPAGE® Antioxidant

Invitrolon™ or Nitrocellulose Membrane/Filter Paper Sandwiches

Blotting Roller

Incubation Tray
Note: Use the blotting pads that fit the XCell II™ Blot Module. Do not use the
E-PAGE Blotting Pads for semi-wet transfer.
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