Thermo Scientific Pierce G2 User manual

Thermo Scientific Pierce
G2 Fast Blotter
Version 1
May 2013
User Manual

Thermo Scientific
© 2013 Thermo Fisher Scientific Inc. All rights reserved. All trademarks are the property of
Thermo Fisher Scientific Inc.and its subsidiaries. Specifications, terms and pricing are subject
to change. Not all products are available in allcountries. Please consult your local sales
representative for details.

Thermo Scientific / Thermo Scientific Pierce G2 Fast Blotter |3
Contents
Section 1 .......................................................................................................................5
Overview .......................................................................................................................5
Contacting Thermo Fisher Scientific............................................................................................5
Instrument Overview....................................................................................................................7
Unpacking andSetup Instructions......................................................................................7
User Interface.....................................................................................................................7
Section 2 .......................................................................................................................9
Instrument.....................................................................................................................9
Installation Guidelines..................................................................................................................9
Space Requirements..........................................................................................................9
Electrical Requirements...................................................................................................10
Environmental Requirements...........................................................................................11
Blotter Images with Feature Locations.............................................................................12
Start Up............................................................................................................................14
Shut Down........................................................................................................................14
Section 3 .....................................................................................................................15
Pre-Programmed Transfer Methods..........................................................................15
Fast Blotting: Constant Amps/Surface Area..............................................................................15
Pierce 1-Step Transfer Buffer....................................................................................................17
Traditional Semi-Dry Transfer Method.......................................................................................20
Section 4 .....................................................................................................................23
Custom Methods ........................................................................................................23
Modifying Pre-Programmed Methods........................................................................................23
Creating a New Custom Method................................................................................................25
Recommended Maximum Watts/Hour Limit..............................................................................27
Section 5 .....................................................................................................................28
Maintenance................................................................................................................28
Cleaning the Blotter LCD Touchscreen .....................................................................................28
Cleaning the Blotter Housing and the Cassette.........................................................................28
Cleaning the Anode and Cathode Plates...................................................................................29
Troubleshooting.........................................................................................................................30

Thermo Scientific / Thermo ScientificPierce G2 Fast Blotter |5
1
Overview
The Thermo Scientific™ Pierce™ G2 Fast Blotter is designed for rapid semi-dry transfer of
proteins frompolyacrylamide gels to nitrocellulose or PVDF membranes. Traditional Western
blotting techniques require atransfer ofone hour to overnight to achieve good transfer
efficiency. Whenused inconjunction with Thermo Scientific™ Pierce™ 1-Step Transfer Buffer,
the Pierce G2 Fast Blotter is designed to provide transfer efficiency in 5-10 minutes that is
equivalent to, or better than, traditional blotting techniques without the need for gel pre-
equilibration. This significant reduction inprotein transfer time is accomplished by optimizing the
ionic strength of the transfer buffer and increasing the current [amps (A)/cm2] flowing through
the transfer stack. The system has been verified to work withcommonly used pre-cast and
homemade SDS-PAGE gels. The PierceG2 Fast Blotter can alsobe used forstandard semi-
dry transfer protocols with Towbin buffer.
Contacting Thermo Fisher Scientific
For technical or service assistance on the Pierce G2 Fast Blotter:
• U.S. Customers: 1-800-874-3723
Fax: [phone]
• International Customers: 1-815-968-0747
Fax: [phone]
SECTION 1

Section 1|Overview
6|/ ThermoScientific Pierce G2 Fast Blotter Thermo Scientific
Technical Support email for U.S. and international customers outside Europe:
[e-mail]
Technical Support email for European customers:
[e-mail]
For more information about the Thermo Scientific ProteinBiology product line,
please visit thermoscientific.com/pierce

Overview |Section 1
Thermo Scientific / Thermo Scientific Pierce G2 Fast Blotter |7
Instrument Overview
Unpacking and Setup Instructions
Unpack your new Pierce G2 Fast Blotter Control Unit and/or Cassette.
Verify that allof the accessories listed below are includedwith your blotter package. Please
note that the control unit and the cassette can be purchased separately or combined.
Your blotterpurchase includes the following accessories (each quantity1):
Included Accessories (Control unit)
Pierce G2 Fast Blotter ControlUnit
Quick Start Guide
Blot Roller
Power Cord with C13 Connector
Included Accessories (Cassette)
Pierce G2 Fast Blotter Cassette
User Interface
The user interface consists ofa color LCDmenu touchscreen on the top of the control unit.
From the Main Menu, five selections are available. Touching the lower right Down arrow allows
you to see the last two methods and the Setup button.

Section 1|Overview
8|/ ThermoScientific Pierce G2 Fast Blotter Thermo Scientific
1. Pre-Programmed Methods This feature allows you toquickly access the recommended
amperage, voltage and time for the size and number of gels that you intend to
simultaneously transfer. The blotter is only intended for usewith the following
consumables:
•Pierce 1-Step Transfer Buffer
•Bio-Rad™ Trans-Blot™ Turbo™ Transfer Stacks
•Towbin Transfer Buffer for Standard Semi-Dry Transfer (only)
2. Recent Methods Access to recently run methods listed from most to least recent.
3. Custom Methods Create, run and save a custom transfer method.
4. Setup Control audio settings, view software and hardware versions, and install software
updates to the user interface.
5. Shutdown Safely shuts down the device.

Thermo Scientific / Thermo Scientific Pierce G2 Fast Blotter |9
2
Instrument
Installation Guidelines
IMPORTANT Review and implement guidelines for proper set-up before installation. Locate
the imager awayfrom water, solvents, corrosive materials,strong magnetic fields and vibration
sources.
Space Requirements
The blotter requires a stable laboratory bench or table. Provide a minimum clearance of
3 inches (7.62cm)at the rear of the blotter toallow for adequate ventilation for cooling and
access to the power cord andswitch. In addition, allowsufficient space at the front of the blotter
for docking and removing thecassette with/from the control unit.
Table 2.1. Control unit dimensions.
Dimensions
Inches
Centimeters
Width
6.44
16.4
Depth
10.88
27.6
Height
6.06
15.40
Table 2.2. Cassette dimensions.
Dimensions
Inches
Centimeters
Width
10.13
25.7
Depth
10.94
27.8
Height
1.38
3.5
SECTION 2

Section 2|Instrument
10 |/ Thermo Scientific Pierce G2 Fast Blotter Thermo Scientific
Electrical Requirements
Note A grounded circuit capable of delivering the appropriate current and voltage is required
for installation. Electrical requirements can be located on the rear panel of the blotter. The
blotter electrical system will adjust to the proper voltage for the respective country. Connect the
power cord to the rear left side panel of the device and plug into a grounded power outlet.
WARNING After running at high current, the anode and cathode plates can become hot.Use
caution when separating the gels and stacks from the plates.When continuously processing
multiple samples at ~5A for a maximum of 2 hours, allow the cassette tocool for 30minutes or
use multiple cassettes to avoid excessive cassette heating.
WARNING Blotter use outside of the workflows described in thismanual may put the operator
at risk of dangerous exposureto electrical shock. Do not use this instrument for any purposes
or in any configurations not described in thismanual.
WARNING The blotter control unit can contain dangerous electricity and is not designed to be
opened by theuser. Disconnect all power to the control unit before maintenance by a qualified
technician.
WARNING Do not overfill thecassette withliquid. Excess liquid can overflow into the control
unit and possibly cause electric shock. Follow the appropriate instructions for reagent amounts
and empty any remaining liquid in the cassetteupon run completion.
Table 2.3. Blotter electrical ratings.
Electrical Parameter Rating
Supply Voltage (VAC)
120-240
Frequency (Hz)
50-60
Power Rating (W)
120
Fuse (power center)
T4AL,250V, 4A

Instrument |Section 2
Thermo Scientific / Thermo Scientific Pierce G2 Fast Blotter |11
Table 2.4. Blotter electrical requirements for major countries.
Region
Voltage (VAC)
Frequency (Hz)
Amps (A)
US and Canada
120
60
1.0
Europe
220
50
0.5
UK
240
50
0.5
Japan
100
50
1.2
China
220
50
0.5
India
230
50
0.5
South Korea
220
60
0.5
Environmental Requirements
For optimal performance, maintain a constant laboratory temperature range of 15-25°C (59-
77°F) with relative humidity between 20-70%. Do not place blotter in direct sunlight.

Section 2|Instrument
12 |/ Thermo Scientific Pierce G2 Fast Blotter Thermo Scientific
Blotter Images with Feature Locations
Figure 2.1. Blotter Control Unit front and side views detailing main components of control unit.
Figure 2.2. Cassette view detailing the upper cathode, the lower anode and the electrical contacts that insert
into the control unit contacts.
Cassette Guide Rails
Color LCD Touchscreen
Control Unit Contact Points
Cassette Contact Points
Anode
Cathode

Instrument |Section 2
Thermo Scientific / Thermo Scientific Pierce G2 Fast Blotter|13
Figure 2.3. Contol unit with cassette inserted.
Figure 2.4. Rear view of control unit with cassette inserted.
Cassette
USB
Cooling Fan
Power Cord
Connector
Power Switch
Control
Unit

Section 2|Instrument
14 |/ Thermo Scientific Pierce G2 Fast Blotter Thermo Scientific
Start Up
Turn the powerswitch located at the bottom rear of the blotter panel to the On position. The
power switch provides power to all of the blottercomponents. The on-board computer boots up,
the touchscreen displays the Thermo Scientific logo followed by “Welcome to the Pierce G2
Fast Blotter,”and then automatically moves to the Main Menu. The blotter is now ready for
operation.
Shut Down
On the Main Menu screen, touch the Shut Down button. When prompted “Areyou sure you
want to shut down?” select Yes. When notified that “It is nowsafe to turn off the device,” turn
the power switch to the Off position.

Pre-Programmed Transfer Methods |Section 3
Thermo Scientific / Thermo Scientific Pierce G2 Fast Blotter |15
3
Pre-Programmed Transfer Methods
Fast Blotting: Constant Amps/Surface Area
Fast blotting methods, such asthe Pierce G2Fast Blotter, require a high-current power supply
and an optimized high ionic strength transfer buffer, such as Pierce 1-Step Transfer Buffer. By
increasing the current, excellent transfer efficiency can be achieved in a much shorter time.
Amperage is held at a constant rate based on the surface area of the transfer stack(s)
(~22-23mA/cm2) and voltage is limited to a maximum of 25V.
Figure 3.1. Fast blotting protocol (volts vs amps).
0.00
0.20
0.40
0.60
0.80
1.00
1.20
0
5
10
15
20
25
0 2 4 6 8 10
Amps
Volts
Time (minutes)
Pierce G2 Fast Blotter: 1 mini gel, 1.3A constant,
25V limit
Volts
Amps
SECTION 3

Section 3|Pre-Programmed Transfer Methods
16 |/ Thermo Scientific Pierce G2 Fast Blotter Thermo Scientific
Traditional, efficient semi-dry transfer protocols use constant voltage (25V) and low-ionic
strength transfer buffer (Towbin) to generate low amperage for an extended length of time
(30-60 minutes).
Figure 3.2. Traditional semi-dry blotting protocol (volts vs amps).
0.00
0.20
0.40
0.60
0.80
1.00
1.20
0
5
10
15
20
25
0 20 40 60
Amps
Volts
Time (minutes)
Traditional Semi-Dry: 1 mini gel, 25V constant,
1.0A limit
Volts
Amps

Pre-Programmed Transfer Methods |Section 3
Thermo Scientific / Thermo Scientific Pierce G2 Fast Blotter |17
Pierce 1-Step Transfer Buffer
Simultaneously transfer one to four mini-sized gels or one to two midi-sized gels in
5-10 minutes.Gels simultaneously transferred must have the same formulation. Mini gels are
defined as ~46cm2and 60cm2. Midi gels are defined as ~110cm2. Note that these dimensions
are the surfacearea of the gel with the gel fingers removed (not of the cassette).
1. Cut four sheets of ~0.83mm thick Western blotting filter paper and one sheet of
nitrocelluloseor PVDFmembrane to the same size as thegel(s).
Note The transfer stack should contain two sheets of ~0.83mm thick filter paper on the
bottom (anode), followed bymembrane,gel and two sheets of ~0.83mm thick Western
blotting filter paper on top. If 0.83mm thick filter paper is notavailable, do not exceed
1.8mm total filter paper thickness for the bottom of the stack or 1.8mm total filter paper
thickness for the top of the stack. Use the appropriate Thermo Scientific filter papers:
Product No. 84873, 0.83mm thick, 7.0cm × 8.4cm (mini-size)
Product No. 84874, 0.83mm thick, 8.0cm × 13.5cm (midi-size)
Product No. 88600, 0.83mm thick, 8.0cm × 10.5cm
2. Equilibrate filter paper and membrane in Pierce 1-Step Transfer Buffer for a minimum of
5 minutes. Use sufficient buffer to completely cover the filterpaper and membrane.
Note PVDF membrane must be wetted with methanol or ethanol before equilibration in
Pierce 1-Step Transfer Buffer.After electrophoresis, remove gel from cassette(s) and
briefly place ina tray containing deionized water or transfer buffer. This will ensure even
wetting, facilitate proper gel placement and improve contactwith the membrane.
3. Assemble stack centered on the bottom half of thecassette (anode) as depicted in Figure
3.3. Use a blot roller to remove any trapped air bubbles.
Note Removal oftrapped air bubbles isessential for high-quality transfer. Two firm passes
with the included blot roller is typically sufficient.
Note If assemblingmore thanone stackon the anodesurface, evenly space and center
the sandwiches so the top cathode surface applies even pressure to the surface areas of
the stack(s). Ensure there is a1cm space around all stack edges to allow any gases to
escape during transfer.

Section 3|Pre-Programmed Transfer Methods
18 |/ Thermo Scientific Pierce G2 Fast Blotter Thermo Scientific
Figure 3-3. Orientation of filter paper, membrane and gel in transfer stack.
4. Gently press down top of cassette (cathode) tolock into place.
5. Slide cassette into the control unit.
6. Select Pre-Programmed Methods in the Main Menu.
7. Using the touchpad, select the number of gels and gelsize (mini ormidi) you wish to
transfer:
2 sheets of pre-wet filter paper
(combined thickness not to
exceed 1.8mm)
Membrane
Anode (+)
Cathode (–)
2 sheets of pre-wet filter paper
(combined thickness not to
exceed 1.8mm)
Gel

Pre-Programmed Transfer Methods |Section 3
Thermo Scientific / Thermo Scientific Pierce G2 Fast Blotter |19
8. Choose the appropriate program to run (constant parameter in box):
a. Low MW (< 25kDa) 25V Amps 5:00 min
b. Mixed Range MW(25-150kDa) 25V Amps 7:00 min
c. High MW (> 150kDa) 25V Amps 10:00 min
d. Std Semi-Dry 25V 1.0A 60:00 min
e. 1.5mm thick gels or unknown size gels 25V Amps 10:00 min
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 extremelyhigh molecular-weight proteins
or for slow-transferring gels. Do not use the StdSemi-Dry transfer program (d) with Pierce
1-Step Transfer Buffer.
9. Select the Start button to begin transfer.
10. Upon transfer completion, remove the transfer stack from the cassette(s) and thoroughly
rinse the topand bottomsection of the cassette.
Note Build-upof buffersalts will reduce cassette function and prevent the cassettefrom
properly opening and closing.Rinse cassette(s) after every use.
WARNING After running at high current, the anode and cathode plates can become hot.Use
caution when separating the gels and stacks from the plates.When continuously processing
multiple samples at ~5A for a maximum of 2 hours, allow the cassette tocool for 30 minutes or
use multiple cassettes to avoid excessive cassette heating.

Section 3|Pre-Programmed Transfer Methods
20 |/ Thermo Scientific Pierce G2 Fast Blotter Thermo Scientific
Traditional Semi-Dry Transfer Method
Simultaneously transfer one to four mini-sized gels or one to two midi-sized gels in
45-60 minutes. Gels simultaneously transferred must have the same formulation.
1. Equilibrate gel(s) for 15 minutes in Towbin transfer buffer (25mM Tris, 192mM glycine,
20% methanol).
2. Cut two extra thick (~2.48mm thick) sheets of Westernblotting filter paper and one sheet of
nitrocelluloseor PVDFmembrane to the same size as gel(s).
Note The transferstack should contain one sheet of ~2.48mm thick filter paper on the
bottom (anode), followed bymembrane,gel and one sheet of~2.48mm thick Western
blotting paper on top. Do not exceed 2.48mm total filter paper thickness for the bottom of
the stack or 2.48mm total filterpaper thickness for the top ofthe stack. Use the appropriate
Thermo Scientific filter paper:
Product No. 88605, 2.48mm thick, 7.0cm × 8.4cm (mini-size)
Product No. 88610, 2.48mm thick, 8.5cm × 9.0cm
Product No. 88615, 2.48mm thick, 8.0cm × 13.5cm (midi-size)
Product No. 88620, 2.48mm thick, 20.0cm × 20.0cm
3. Equilibrate filter paper and membrane in Towbin transfer buffer for at least 5 minutes.
Note PVDF membrane must be wetted with methanol or ethanol before equilibration in
Towbin transfer buffer.
Note Filter paper and membrane stacks can be prepared ahead (few hours to overnight)
and stored at 4°C.
4. Assemble stack on anode as depicted in Figure 3.4. Center stack(s) on anode to ensure
even pressure. Use blot rollerto remove any trapped air bubbles.
Note Removal oftrapped air bubbles isessential for high-quality transfer. Two firm passes
with the included blot roller is typically sufficient.
Note If assemblingmore thanone stackon the anodesurface, evenly space and center
the sandwiches so the top cathode surface applies even pressure to the surface areas of
the stack(s). Ensure there is a1cm space around all stack edges to allow any gases to
escape during transfer.