Select the appropriate program to run:
a.
Low MW (< 25kDa)
b.
Mixed Range MW (25-150kDa)
c.
High MW( >150kDa)
d.
Std Semi-Dry
e.
1.5mm thick gels or if unknown
Note:
For fast-blotting programs (a, b, c and e), use Pierce
1-Step Transfer Buffer. Transfer time may be increased to
12 minutes for extremely high molecular-weight proteins. Do not
use the Std Semi-Dry transfer program (d) with Pierce 1-Step
Transfer Buffer.
Select the
Start
button to begin transfer.
Pierce 1-Step Transfer Buffer is a highly concentrated salt solution.
Thoroughly wash the anode and cathode after each use and rinse
the unassembled cassette under hot water while removing any
sticky salt residue with a gloved hand. Rinse with deionized water
and stand parts in a rack to dry. For more thorough cleaning, im-
merse the cassette top (cathode) and bottom (anode) in hot water
and use a gloved hand or clean sponge to remove salt residue.
Rinse with deionized water and stand parts in a rack to dry.
NOTE: Failure to keep cassette top and bottom clean can
result in moving parts sticking and lead to poor transfer
efficiency.
After running at high current, the anode and cathode plates
can become hot. Use caution when separating the gels and
stacks from the plates.
IMPORTANT
Review and implement guidelines for proper set-up
before installation. Gels simultaneously transferred must have the
same formulation.
Using the appropriate power cord supplied with the blotter,
connect the blotter control unit to an outlet.
Switch on the Power Button located on the control unit’s rear panel.
For each gel, use four sheets of ~0.83mm thick Western blotting
filter paper and one sheet of nitrocellulose or PVDF membrane cut
to the same size.
Equilibrate filter paper and membrane in Thermo Scientific
™
Pierce
™
1-Step Transfer Buffer for at least 5 minutes. Use sufficient buffer
to cover filter paper and membrane [~50mL per mini-sized (7cm
x 8.4cm) sandwich and ~100mL per midi-sized (8cm x 13.5cm)
sandwich].
Note:
PVDF membrane must be wetted with methanol or
ethanol before equilibration in Pierce 1-Step Transfer Buffer.
After electrophoresis, remove gel(s) from cassette(s) and briefly
place into tray containing deionized water or transfer buffer. This will
ensure even wetting, facilitate proper gel placement and improve
gel contact with the membrane.
Assemble and center sandwich(es) on bottom of cassette (anode)
to ensure even pressure (10mm spaces between sandwiches).
Use a blot roller to remove any trapped air bubbles.
1
2
3
4
Cathode (-)
2 sheets of pre-wet filter paper (combined
thickness not to exceed 1.8mm)
Gel
Membrane
2 sheets of pre-wet filter paper (combined
thickness not to exceed 1.8mm)
Anode (+)
Lock top of the cassette (cathode) into place and slide cassette
into the control unit.
Select
Pre-Programmed Methods
in the Main Menu.
5
6
7
8
10
Quick Start Guide
Thermo Scientific Pierce G2 Fast Blotter
11
Select the number of gels and gel size (mini or midi) for transfer.
a.
1 mini-gel
b.
2 mini-gels or 1 midi-gel
c.
3 mini-gels
d.
4 mini-gels or 2 midi-gels
9
12
