C.B.S. Scientific DCX-800 - Manual (Page 15)

15 www.cbsscientific.com
SECTION 5
Instructions for Western Blotting
5.1 Preparing the Unit for Blotting
1. Remove safety cover from the assembled unit
by simultaneously pressing down on white push
pins while lifting up on blue safety cover.
Do not
remove safety cover by pulling up on leads!
Remove white core from lower reservoir by grasp-
ing core with one hand and lifting directly up.
Open doors on the core assembly by pulling up
on the white latches, as shown in figure 1.
2. Open blotting cassette as shown in figures 2-3
and lay it flat on the bench.
3. Assemble blotting stack as shown in figure 4. With
cassette wide open assemble components on
black side in the following order: foam pad, gel*,
buffer saturated transfer membrane, then buf-
fer saturated blotting paper. Smooth with gloved
finger or roll with glass rod to be sure no bubbles
exist between the gel an the transfer membrane.
*Note: to prepare gel for blotting, trim off wells
and any excess acrylamide at the bottom, and
invert 180º so that the large molecular weight
proteins are at the bottom of the cassette. This
puts them in contact with a stronger field strength
and allows the blotting transfer to take place more
efficiently.
red side
high molecular weight
bands at this end
blotting paper
gel
foam pad
black side
membrane
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SECTION 4
Alternate Protocol for Slab Gel Electrophoresis.
(Not using Pre-cast gels)
4.2 Preparing the Electrophoresis Unit when Using Gel Wrap™ Gasket
Casting Method
1. Place unit in authorized work area. Remove
safety cover from the assembled unit by simul-
taneously pressing down on white push pins
while lifting up on blue safety cover as shown in
figure 1.
Do not remove safety cover by pulling up on
leads!
2. Remove white core from lower reservoir by
grasping core with one hand and lifting directly
up as shown in figure 2.
3. Open doors on the core assembly by pulling up
on the white latches, as shown in figure 3.
4. Slide glass plate sandwich into the core assem-
bly with the notched plate facing in towards the
upper buffer reservoir.
5. If running one gel, slide white plastic adaptor
plate into the side without the gel.
6. Close doors and relatch by pressing down on
the white latches so that the assembly looks like
that shown in figure 4.
7. Place stirring bar into bottom of reservoir in stir-
ring corral (as shown in figure 7 on page 10).
8. If using cooling capability of DCX-800C attach
external cooler to tubing adapters and begin
circulating coolant through labyrinth.
9. For rest of protocol please refer to instructions
on pages 10-11 (Sections 3.2 and 3.3) for run-
ning the gel and removing it after electrophore-
sis.
Go to Section 3.2
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