37
Semi-Wet Blotting of E-PAGE
™
Gels,
Continued
Semi-Wet
Blotting
Procedure
1.
Remove any small pieces of gel from the well areas by gently rubbing a
gloved finger over the well side of the gel. The gel pieces on the gel may
cause air bubbles and field distortion during transfer.
2.
Lay the gel on a flat surface, well side up, in a suitable container and pour
sufficient 1X NuPAGE
®
Transfer Buffer (see previous page) over the gel to
fill all wells.
3.
Place a piece of pre-soaked filter paper on top of the gel and remove any
trapped air bubbles by gently rolling the Blotting Roller over the filter
paper. Turn the gel over so the filter paper and gel and are facing
downwards over a gloved hand or clean flat surface
4.
Place a pre-soaked membrane on the gel and remove any trapped air
bubbles with the Blotting Roller.
5.
Place another pre-soaked filter paper on top of the membrane and remove
any trapped air bubbles with the Blotting Roller.
6.
Place two pre-soaked blotting pads into the cathode core of the blot module.
Note:
Use the blotting pads that fit the XCell II
™
Blot Module.
Do not
use
the E-PAGE
Blotting Pads for semi-wet transfer.
7.
Place the gel/membrane assembly on the blotting pads, such that the gel is
closest to the cathode core.
8.
Add another pre-soaked blotting pad on top of the membrane assembly.
Remove air bubbles with the Blotting Roller.
9.
Add enough pre-soaked blotting pads to rise to 0.5 cm over the rim of the
cathode core. Place the anode (+) core on top of the pads.
10.
Hold the blot module together firmly and slide it into the guide rails on the
lower buffer chamber.
11.
Insert the gel tension wedge into the lower buffer chamber and lock the
wedge into position.
12.
Fill the blot module with 1X NuPAGE
®
Transfer Buffer (see page 36) until
the gel/membrane assembly is just covered.
13.
Fill the outer buffer chamber with 650 mL deionized water.
14.
Place the lid on the unit and connect electrical leads to the power supply.
15.
Perform transfer for nitrocellulose or PVDF membranes using 35 V constant
for 1 hour. The expected starting current is about 170 mA.
Summary of Contents for E-PAGE Gels
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