Thermo Scientific Pierce G2 User Manual

Thermo Scientific Pierce

User Manual

G2 Fast Blotter

Version 1

May 2013

© 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 all countries. Please consult your local sales
representative for details.

Thermo Scie ntific

Contents

Section 1 ....................................................................................................................... 5

Overview ....................................................................................................................... 5

Contacting Thermo Fisher Scientific ............................................................................................ 5

Instrument Overview .................................................................................................................... 7

Unpacking and Setup 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 Scientific Pierce G2 Fast Blotter | 3

1

SECTION 1

Overview

The Thermo Scientific™ Pierce™ G2 Fast Blotter is designed for rapid semi-dry transfer of
proteins from polyacrylamide gels to nitrocellulose or PVDF membranes. Traditional Western
blotting techniques require a transfer of one hour to overnight to achieve good transfer
efficiency. When used in conjunction 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 pre­equilibration. This significant reduction in protein transfer time is accomplished by optimizing the

2

ionic strength of the transfer buffer and increasing the current [amps (A)/cm
the transfer stack. The system has been verified to work with commonly used pre-cast and
homemade SDS-PAGE gels. The Pierce G2 Fast Blotter can also be used for standard semi­dry transfer protocols with Towbin buffer.

Contacting Thermo Fisher Scientifi c

] flowing through

For technical or service assistance on the Pierce G2 Fast Blotter:

• U.S. Customers: 1-800-874-3723

• International Customers: 1-815-968-0747

Thermo Scie ntific / T hermo Scienti fic Pierce G2 Fast Blotter | 5

Fax: [phone]

Fax: [phone]

Section 1 | Overview

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 Protein Biology product line,

please visit thermoscientific.com/pierce

6 | / Thermo Scient if ic Pierc e G2 Fas t Blot ter Thermo Scie ntific

Overview | Section 1

Instrument Overview

Unpacking and Setup Instructions

Unpack your new Pierce G2 Fast Blotter Control Unit and/or Cassette.

Verify that all of the accessories listed below are included with your blotter package. Please
note that the control unit and the cassette can be purchased separately or combined.

Your blotter purchase includes the following accessories (each quantity 1):

Included Accessories (Control unit)

Pierce G2 Fast Blotter Control Unit
Quick Start Guide
Blot Roller
Power Cord with C13 Connector

Included Accessories (Cassette)

Pierce G2 Fast Blotter Cassette

User Interface

The user interface consists of a color LCD menu 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.

Thermo Scientific / Thermo Scientific Pierce G2 Fast Blotter | 7

Section 1 | Overview

1. Pre-Programmed Methods This feature allows you to quickly 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 use with 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 version s, and install software
updates to the user interface.

5. Shutdown Safely shuts down the device.

8 | / Thermo Scient if ic Pierc e G2 Fas t Blot ter Thermo Scie ntific

2

Dimensions

Inches

Centimeters

Width

6.44

16.4

Depth

10.88

27.6

Height

6.06

15.40

Dimensions

Inches

Centimeters

Width

10.13

25.7

Depth

10.94

27.8

Height

1.38

3.5

SECTION 2

Instrument

Installation Guidelines

IMPORTANT Review and implement guidelines for proper set-up before installation. Locate
the imager away from 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 to allow for adequate ventilation for cooling and
access to the power cord and switch. In addition, allow sufficient space at the front of the blotter
for docking and removing the cassette with/from the control unit.

Table 2.1. Control unit dimensions.

Table 2.2. Cassette dimensions.

Thermo Scientific / Thermo Scientific Pierce G2 Fast Blotter | 9

Section 2 | Instrument

Supply Voltage (VAC)

120-240

Frequency (Hz)

50-60

Power Rating ( W)

120

Fuse (power center)

T4AL,250V, 4A

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 to cool for 30 minutes or
use multiple cassettes to avoid excessive cassette heating.

WARNING Blotter use outside of the workflows described in this manual may put the operator
at risk of dangerous exposure to electrical shock. Do not use this instrument for any purposes
or in any configurations not described in this manual.

WARNING The blotter control unit can contain dangerous electricity and is not designed to be
opened by the user. Disconnect all power to the control unit before maintenance by a qualified
technician.

WARNING Do not overfill the cassette with liquid. 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 cassette upon run completion.

Table 2.3. Blotter electrical ratings.

Electrical Parameter Rating

10 | / Thermo Scientific Pierce G2 Fast Blot ter Thermo Scie ntific

Instrument | Section 2

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

Table 2.4. Blotter electrical requirements for major countries.

Environmental Requirements

For optimal performance, maintain a constant laboratory temperature range of 15-25°C (59­77°F) with relative humidity between 20-70%. Do not place blotter in direct sunlight.

Thermo Scientific / Thermo Scientific Pierce G2 Fast Blotter | 11

Section 2 | Instrument

Cassette Guide Rails

Color LCD Touchscreen

Control Unit Contact Points

Cassette Contact Points

Anode

Cathode

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 l ower anode and the electrical contacts that insert
into the control unit contacts.

12 | / Thermo Scientific Pierce G2 Fast Blot ter Thermo Scie ntific

Cassette

USB

Cooling Fan

Power Cord

Power Switch

Control

Unit

Figure 2.3. Contol unit with cassette inserted.

Instrument | Section 2

Figure 2.4. Rear view of control unit with cassette inserted.

Thermo Scientific / Thermo Scientific Pierce G2 Fast Blotter | 13

Connector

Section 2 | Instrument

Start Up

Turn the power switch located at the bottom rear of the blotter panel to the On position. The
power switch provides power to all of the blotter components. 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 “Are you sure you
want to shut down?” select Yes. When notified that “It is now safe to turn off the device,” turn
the power switch to the Off position.

14 | / Thermo Scientific Pierce G2 Fast Blot ter Thermo Scie ntific

Pre-Programmed Transfer Methods | Section 3

Amps

Pierce G2 Fast Blott er: 1 min i gel, 1.3A constant,

Volts

Amps

SECTION 3

3

Pre-Programmed Transfer Methods

Fast Blotting: Constant Amps/S ur face Area

Fast blotting methods, such as the Pierce G2 Fast 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)

2

(~22-23mA/cm

25
20
15

Volts

10

Figure 3.1. Fast blotting protocol (volts vs am ps).

) and voltage is limited to a maxi mum of 25V.

5
0

0 2 4 6 8 10

25V limit

1.20

1.00

0.80

0.60

0.40

0.20

0.00

Time (minutes)

Thermo Scientific / Thermo Scientific Pierce G2 Fast Blotter | 15

Section 3 | Pre-Programmed Transf er Meth o ds

Volts

Amps

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).

Traditional Semi-Dry : 1 mini gel, 25V const ant ,

1.0A limit

25
20
15

Volts

10

5
0

0 20 40 60

Time (minutes)

Figure 3.2. Traditional semi-dry blotting protocol (volts vs amps).

1.20

1.00

0.80

0.60

0.40

0.20

0.00

Amps

16 | / Thermo Scientific Pierce G2 Fast Blot ter Thermo Scie ntific

Pre-Programmed Transfer Methods | Section 3

Pierce 1-Step Transfer B uffer

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

2

defined as ~46cm
are the surface area 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
nitrocellulose or PVDF membrane to the same size as the gel(s).

Note The transfer stack should contain two sheets of ~0.83mm thick filter paper on the
bottom (anode), followed by membrane, gel and two sheets of ~0.83mm thick Western
blotting filter paper on top. If 0.83mm thick filter paper is not available, 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

and 60cm2. Midi gels are defined as ~110cm2. Note that these dimensions

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 filter paper 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 in a tray containing deionized water or transfer buffer. This will ensure even
wetting, facilitate proper gel placement and improve contact with the membrane.

3. Assemble stack centered on the bottom half of the cassette (anode) as depicted in Figure

3.3. Use a blot roller to remove any trapped air bubbles.

Note Removal of trapped air bubbles is essential for high-quality transfer. Two firm passes
with the included blot roller is typically sufficient.

Note If assembling more than one stack on the anode surface, 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 a 1cm space around all stack edges to allow any gases to
escape during transfer.

Thermo Scientific / Thermo Scientific Pierce G2 Fast Blotter | 17

Section 3 | Pre-Programmed Transf er Meth o ds

2 sheets of pre-wet filter paper
exceed 1.8mm)

Membrane

Anode (+)

Cathode (–)

2 sheets of pre-wet filter paper

Gel

(combined thickness not to

(combined thickness not to
exceed 1.8mm)

Figure 3-3. Orientation of filter paper, membrane and gel in transfer stack.

4. Gently press down top of cassette (cathode) to lock 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 gel size (mini or midi) you wish to
transfer:

18 | / Thermo Scientific Pierce G2 Fast Blot ter Thermo Scie ntific

Pre-Programmed Transfer Methods | Section 3

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 extremely high molecular-weight proteins
or for slow-transferring gels. Do not use the Std Semi-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 top and bottom section of the cassette.

Note Build-up of buffer salts will reduce cassette function and prevent the cassette from
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 to cool for 30 minutes or
use multiple cassettes to avoid excessive cassette heating.

Thermo Scientific / Thermo Scientific Pierce G2 Fast Blotter | 19

Section 3 | Pre-Programmed Transf er Meth o ds

Traditional Se mi-Dry Transfer M ethod

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 Western blotting filter paper and one sheet of
nitrocellulose or PVDF membrane to the same size as gel(s).

Note The transfer stack should contain one sheet of ~2.48mm thick filter paper on the
bottom (anode), followed by membrane, 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 filter paper thickness for the top of the 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 roller to remove any trapped air bubbles.

Note Removal of trapped air bubbles is essential for high-quality transfer. Two firm passes
with the included blot roller is typically sufficient.

Note If assembling more than one stack on the anode surface, 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 a 1cm space around all stack edges to allow any gases to
escape during transfer.

20 | / Thermo Scientific Pierce G2 Fast Blotter Thermo Scientific

Pre-Programmed Transfer Methods | Section 3

1 sheet of pre-wet filter paper

2.48mm)

Membrane

Anode (+)

Cathode (–)

1 sheet of pre-wet filter paper

Gel

(thickness not to exceed

(thickness not to exceed

2.48mm)

Figure 3-4. Orientation of filte r paper, membrane and gel in transfer stack for standard semi-dry protocol.

5. Gently press down top of cassette (cathode) to lock into place. Slide cassette into the
control unit.

6. Select Pre-Programmed Methods in the Main Menu.

7. Select the number of gels and gel size (mini or midi) you wish to transfer:

a. 1 mini-sized gel 25V
b. 2 mini-sized gels or 1 midi-sized gel 25V
c. 3 mini-sized gels 25V
d. 4 mini-sized gels or 2 midi-sized gels 25V

Thermo Scientific / Thermo Scientific Pierce G2 Fast Blotter | 21

Section 3 | Pre-Programmed Transf er Meth o ds

8. Choose program Std Semi-dry (constant parameter in box):

a. Low MW 25V Amps 5:00 min

b. Mixed Range MW 25V Amps 7:00 min

c. High MW 25V Amps 10:00 min

d. Std Semi-dry 25V 1.0A 60:00 min

e. 1.5 mm thick gels or unknown size gels 25V Amps 10:00 min

Note For Std Semi-dry program (d), use Towbin transfer buffer or other conventional transfer
buffer.

9. Select the Start button to begin transfer.

After running at high current, the anode and cathode plates can become hot. Use caution when
separating the gels and stacks from the plates.

22 | / Thermo Scientific Pierce G2 Fast Blot ter Thermo Scie ntific

4

SECTION 4

Custom Methods

Custom Methods | Section 4

Modifying Pre-P rogram m ed Met hods

Pre-programmed methods can be modified before Start by selecting the Modify button.

1. Pressing the Select Constant (V or A) button will toggle the constant variable parameter
from amps to volts or volts to amps.

2. Highlight the variable for change and press the Up or Down arrow to raise or lower the
selected variable’s value.

Thermo Scientific / Thermo Scientific Pierce G2 Fast Blotter | 23

Section 4 | Custom Methods

3. After the program is modified, press the Done button.

4. The “Review Method” screen is now displayed. Press Run Without Saving or Save.

a. Run Without Saving will prompt the “Transfer Ready Screen.” Press Start to begin

the modified method.

b. If you wish to save the modified method, press Save and use the alphanumeric key

pad to enter up to 15 characters to identify the new custom method. Press Done and
then Start to begin the method. The custom method will be saved in Custom

Methods in the Main Menu.

24 | / Thermo Scientific Pier ce G2 Fast Blot ter Thermo Scie ntific

Custom Methods | Section 4

Creating a New Cus tom Method

Custom methods are typically used when transferring non-standard gel sizes (i.e., not mini- or
midi-sized). For rapid transfer protocols using Pierce 1-Step Transfer Buffer, measure the total

2

surface area of the gel(s) that you are transferring and multiply by 23mA/cm
transferring a 12cm × 15cm non-standard gel (measuring the gel and not the cassette), then
use you would use the following formula:

2

(12cm × 15cm × 23mA/cm

) ÷ 1000mA/A = 4.1A (constant)

The voltage limit will be set at 25V. The amperage will be constant and set at 4.1A. The time
will be set at 7 minutes for mid-range molecular weight proteins (25-150kDa).

. For example, if

Note Five amps is the maximum current setting allowed. If the gel area exceeds 220cm

2

, set
the amperage to 5A and compensate by increasing the transfer time by 1-2 minutes. For
standard semi-dry protocols using Towbin buffer, voltage is held constant at 25V, amperage is
limited to 1.0A and time is set to 30 minutes to 1 hour. Standard semi-dry programming does
not take surface area into consideration.

1. To create a custom method, press the Custom Method button on the Main Menu.

2. Select the Create Method button and press Select Constant (V or A) to toggle the

constant variable parameter from amps to volts or volts to amps.

3. Highlight the variable for change and press the Up or Down arrow to raise or lower the

Thermo Scientific / Thermo Scientific Pierce G2 Fast Blotter | 25

selected variable’s value.

Section 4 | Custom Methods

4. After the program is modified, press the Done button.

5. The Review Method screen is now displayed. Select Run Without Saving or Save.

a. Run Without Saving will prompt the Transfer Ready screen. Press Start to begin the

modified method.

b. If you wish to save the modified method, press Save and use the alphanumeric

keypad to enter up to 15 characters to identify the new custom method. Press Done
and then Start to begin the method. The custom method will be saved in Custom
Methods in the Main Menu.

26 | / Thermo Scientific Pierce G2 Fast Blot ter Thermo Scie ntific

Custom Methods | Section 4

Recommended Maximum Watts/Hour Limit

To prevent damage to the cassette or control unit, custom methods are limited to
25 watts/hour (when modifying pre-programmed methods and/or creating a new method). Any
voltage or amperage adjustments that result in exceeding the maximum 25 watts/hour will bring
up an alert triangle on the Up arrow key. Continuing past this value will eventual ly result in
a Watt-Hour Limit warning window.

If the Watt-Hour Limit warning window appears, select the Okay button to return to the
previous screen and decrease appropriate number to return to a watt-hour value less than
25 watts/hour.

1. Pierce 1-Step Transfer Buffer is intended to rapidly transfer protein from gel to membrane

using a high transfer current (based on transfer area) and a short time (5-10 minutes).

a. Do not exceed 12-minute transfer times with rapid transfer protocols.

b. Do not exceed 22-23mA/cm

2. Conventional transfer buffers such as Towbin buffer (25mM Tris, 192mM glycine, 20%

methanol) are low ionic strength buffers and transfer time ranges from 45-60 minutes.
Current normally spikes and then quickly drops to a very low level, while voltage reaches
its maximum (25 volts).

2

(current/surface area) with rapid transfer protocols.

Thermo Scientific / Thermo Scientific Pierce G2 Fast Blotter | 27

Section 5 | Maintenance

SECTION 5

5

Maintenance

Cleaning the Blot ter LCD Touchscreen

IMPORTANT Use of ammonia-based cleaners to clean the touchscreen is NOT
recommended. Do not spray cleaner directly onto the LCD screen.

IMPORTANT Do not press hard while cleaning the screen. Do not leave excess cleaning
solution on the LCD screen as this may cause long-term damage.

1. Use a pre-moistened LCD screen-cleaning tissue to clean the surface of the touchscreen.

Cleaning the Blot ter Housing and the Cas sette

IMPORTANT When using cleaning products with the blotter, consult the appropriate material
safety data sheet (MSDS) to confirm compatibility and proper usage of the cleaning product.

1. Spray a soft cloth or paper towel with cleaning solution (e.g., general-purpose cleaner

containing water and mild detergent, Isopropanol, or 70% ethanol). Avoid abrasives and
organic solvents.

2. Wipe the exterior surfaces of the control unit and cassette. Use caution to avoid touching

3. Ensure that the cooling fan vent is free of dust and debris.

28 | / Thermo Scientific Pierce G2 Fast Blotter Thermo Sc ientific

the LCD touchscreen with anything other than a screen-cleaning wipe.

Maintenance | Section 5

Cleaning the Anod e and Cathode Plates

IMPORTANT Always wear gloves when touching the anode and/or cathode.

Note Pierce 1-Step Transfer Buffer is a highly concentrated salt solution. Build-up of buffer

salts will reduce cassette function and prevent the cassette from properly opening and closing.

1. After each use, thoroughly rinse the anode and cathode under warm water while removing

any sticky salt residue with a gloved hand. If the cathode has excessive salt build-up, use a
mild acetic acid solution (< 5%) to remove the residue.

2. Rinse with deionized water and stand parts in a rack to dry.

Thermo Scientific / Thermo Scientific Pierce G2 Fast Blotter | 29

Section 5 | Maintenance

Problem

Possible Cause

Solution

Troubleshooting

Cassette is difficult to
open

Inconsistent transfer

Salt deposited on moving
parts inside cassette

Membrane or filter paper
was insufficiently
equilibrated in Pierce
1-Step Transfer Buffer

Insufficient transfer time Increase transfer time from 7-10 minutes to

Rinse the cassette top and bottom under warm water
while removing any sticky salt residue with a gloved
hand. Briefly rinse with deionized water and place in a
rack to dry. For more thorough cleaning, immerse the
unassembled cassette in warm water, use a gloved
hand or clean sponge to remove any sticky salt
residue. Rinse with deionized water and place
perpendicular in a rack to dry. NOTE: Failure to keep
cassette top (cathode) and bottom (anode) clean will
result in the sticking of moving parts and lead to poor
transfer efficiency

Equilibrate membrane and filter paper in Pierce
1-Step Transfer Buffer before transfer for a minimum
of 5 minutes. Use a sufficient amount of buffer for the
equilibration step

10-12 minutes

Inefficient transfer of
low molecular-weight
proteins to PVDF

PVDF membrane was not
pre-wetted with methanol or
ethanol

Air bubbles trapped
between gel and membrane

Filter paper used in the fast
transfer exceeded 1.8mm

Inefficient binding of some
low molecular-weight
proteins (< 25kDa) to PVDF
membrane

Wet PVDF membrane with methanol or ethanol and
equilibrate for 10-15 minutes in Pierce 1-Step
Transfer Buffer before transfer

When assembling transfer stack, use a roller or
pipette to remove any air bubbles between the gel
and the membrane

Use filter paper < 1.8mm thick

Combine ethanol and Pierce 1-Step Transfer Buffer in
a 15:85 ratio before equilibrating filter paper and
membrane (e.g., 15mL of ethanol with 85mL of Pierce
1-Step Transfer Buffer)

30 | / Thermo Scientific Pierce G2 Fast Blot ter Thermo Scie ntific

2162527.0 05/13 Prin ted in China

thermoscientific.com/pierce

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