35
Semi-Wet Blotting of E-PAGE
™
Gels
Introduction
A semi-wet blotting procedure using an XCell II
™
Blot Module for transfer of
proteins from E-PAGE
™
Gels is described below.
Semi-Wet
Transfer
Apparatus
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 both 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.
Summary of Contents for E-PAGE Gels
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