Thermo Scientific Pierce 22834 Quick Start Manual

Thermo Scientific Pierce Electrophoretic Blotting
Quick Start Guide
Thermo Scientific Pierce
™
or Thermo Scientific Pierce
™
IMPORTANT Review and implement guidelines for proper set-up
before installation. For detailed instructions refer to the manual at
www.thermoscientific.com/pierce-power-system.
1
Using the supplied power cord, connect the Pierce Power Station to an
electrical outlet.
2
Press the Power Button (located at the rear) to turn ON the Pierce
Power Station.
IMPORTANT: For first use, refer to "Using the Pierce Power
Station for First Time" section. If you currently use a Pierce Power
Station for staining and are adding blotting functionality, refer to
"Using the Pierce Power Blot Cassette for First Time" section.
Upgraded Pierce G2 Fast Blotter/Stainer Control Units do not require
any special activation in order to recognize the Power Stain Cassette
or the Power Blot Cassette.
3 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.
4
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.
5
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 membrane.
6
Assemble and center sandwich(es) on the anode (see figure below)
to ensure even pressure. When transferring two gels allow for 10mm
spaces between sandwiches. Use a blot roller to remove any trapped
air bubbles.
7 Lock top of the cassette (cathode) into place and slide the assembled
cassette into the Pierce Power Station.
8
Select Begin Blotting.
Power Blotter (# 22834), Thermo Scientific
™
G2 Fast Blotter (# 62291 upgraded with # 62287), and Thermo Scientific
™
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 (+)
Pierce Power System (# 22830),
™ ™
™
Select Pre-Programmed Methods in the Main Menu.
9
10 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
Select Gel Number and Size
Blot Cassette
11
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 (program a, b, c and e) use
Pierce 1-Step Transfer Buffer. Transfer time may be increased to
12 minutes for high molecular-weight proteins (> 200kDa). Do not
use the Std Semi-Dry transfer program (program d) with Pierce
1-Step Transfer Buffer.
Choose Program
1 mini-gel
Low MW
6 V 1.3 A 5:00
Mid-Range MW
11 V 1.3 A 7:00
High MW
18 V 1.3 A 10:00
12
Select the Start button to begin transfer.
13
Pierce 1-Step Transfer Buffer is a highly concentrated salt solution.
Thoroughly wash the anode and cathode after each use by rinsing
the unassembled cassette under hot water while removing any salt
residue with a gloved hand. Rinse with deionized water and stand
parts in a rack to dry. For more thorough cleaning, immerse the un-
assembled 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 sticking of movable parts and
lead to poor transfer efficiency.
Pierce Power Blot Cassette (# 22835)
™
Back
Back