Invitrogen iBlot 2 Manual de usuario
For Research Use Only. Not for use in diagnostic procedures.
iBlot™2 Dry Blotting System
USER GUIDE
For dry, electroblotting of proteins from mini-, midi-, and E-
PAGE™gels
Catalog Number IB21001
Publication Number MAN0009112
Revision E.0
Life Technologies Corporation | 5781 Van Allen Way | Carlsbad, CA 92008
For descriptions of symbols on product labels or product documents, go to thermofisher.com/symbols-definition.
The information in this guide is subject to change without notice.
DISCLAIMER: TO THE EXTENT ALLOWED BY LAW, THERMO FISHER SCIENTIFIC INC. AND/OR ITS AFFILIATE(S) WILL NOT BE LIABLE FOR SPECIAL,
INCIDENTAL, INDIRECT, PUNITIVE, MULTIPLE, OR CONSEQUENTIAL DAMAGES IN CONNECTION WITH OR ARISING FROM THIS DOCUMENT,
INCLUDING YOUR USE OF IT.
Revision history: Pub. No. MAN0009112
Revision Date Description
E.0 10 January 2019 Replacing pictures in the "Replacing electrical contacts" section.
Converted to CCMS.
D.0 15 March 2017 Addition of detailed information on proper method of closing device lid.
Important Licensing Information: These products may be covered by one or more Limited Use Label Licenses. By use of these products, you accept
the terms and conditions of all applicable Limited Use Label Licenses.
NOTICE TO PURCHASER: DISCLAIMER OF LICENSE: Purchase of this software product alone does not imply any license under any process,
instrument or other apparatus, system, composition, reagent or kit rights under patent claims owned or otherwise controlled by Life Technologies
Corporation, either expressly, or by estoppel.
TRADEMARKS: All trademarks are the property of Thermo Fisher Scientific and its subsidiaries unless otherwise specified.
©2019 Thermo Fisher Scientific Inc. All rights reserved.
Contents
■CHAPTER 1 Product information ....................................... 6
Product contents ................................................................ 6
Types of products ........................................................... 6
iBlot™2 Gel Transfer Device contents .......................................... 6
Before starting ............................................................. 6
Installing the instrument .................................................... 7
Service operation requirements .............................................. 8
Unpacking instructions .......................................................... 8
Upon receiving the instrument ................................................ 8
Unpacking the iBlot™2 Gel Transfer Device .................................... 8
iBlot™2 Gel Transfer Device ...................................................... 8
Front view ................................................................. 8
Rear view .................................................................. 9
Side view .................................................................. 9
About the system ............................................................... 9
iBlot™2 Dry Blotting System ................................................. 9
Features .................................................................. 10
System components ........................................................ 10
System overview ........................................................... 10
Description of parts ............................................................ 12
iBlot™2 Gel Transfer Device ................................................. 12
Blotting roller ............................................................. 13
Stylus .................................................................... 14
Power cord ............................................................... 14
Power adapters ........................................................... 14
iBlot™2 Transfer Stacks .................................................... 15
Guidelines for iBlot™2 Transfer Stacks ....................................... 15
Top stack ................................................................. 16
Bottom stack .............................................................. 16
Transfer membrane ........................................................ 16
iBlot™Filter Paper ......................................................... 17
iBlot™2 absorbent pad ..................................................... 17
Operating the iBlot™2 Gel Transfer Device ........................................ 18
First time usage of the iBlot™2 Gel Transfer Device ............................ 18
Control panel of the iBlot™2 Gel Transfer Device .............................. 18
iBlot™2 Gel Transfer Device options screen ................................... 19
iBlot
™
2 Dry Blotting System User Guide
3
Setting date and time ....................................................... 20
iBlot™2 Gel Transfer Device tutorial ......................................... 20
Writing logs to a USB storage device ......................................... 21
iBlot™2 Gel Transfer Device application notes ................................. 21
Upgrading iBlot™2 Gel Transfer Device firmware .............................. 22
Resetting the iBlot™2 Gel Transfer Device to factory settings .................... 22
Description of methods ......................................................... 23
Methods .................................................................. 23
Recommended running parameters .......................................... 23
■CHAPTER 2 Protein transfer protocol ................................ 25
Experimental overview ......................................................... 25
Experimental outline ....................................................... 25
General guidelines ............................................................. 25
Recommended gel types .................................................... 26
Using the iBlot™2 Gel Transfer Device ............................................ 26
Introduction ............................................................... 26
Materials needed .......................................................... 26
Selecting a method ........................................................ 27
Removing the gel .......................................................... 28
General guidelines ......................................................... 28
Assembling the iBlot™2 Transfer Stack ....................................... 29
Performing blotting ........................................................ 33
Disassembling the iBlot™2 Transfer Stack ........................................ 34
Introduction ............................................................... 34
Procedure ................................................................ 34
■CHAPTER 3 Custom methods .......................................... 36
Creating custom methods ....................................................... 36
Introduction ............................................................... 36
Programming a custom method ............................................. 36
Save a custom method ..................................................... 38
Creating custom methods from a template ........................................ 39
Introduction ............................................................... 39
Select a template .......................................................... 39
Change voltage/time ....................................................... 40
Add/remove steps ......................................................... 41
Save a custom method ..................................................... 43
■APPENDIX A Troubleshooting ......................................... 45
Introduction ................................................................... 45
Contents
4
iBlot
™
2 Dry Blotting System User Guide
■APPENDIX B Post transfer analysis ................................... 48
Post transfer analysis .......................................................... 48
■APPENDIX C Optimizing blotting ...................................... 49
Optimizing blotting ............................................................. 49
■APPENDIX D Maintenance ............................................. 50
Cleaning ...................................................................... 50
General maintenance ........................................................... 50
Replacing the fuse ............................................................. 50
Replacing electrical contacts .................................................... 51
■APPENDIX E Product specifications .................................. 55
iBlot™2 Gel Transfer Device specifications ........................................ 55
iBlot™2 Transfer Stack specifications ............................................. 55
■APPENDIX F Accessory products ...................................... 57
iBlot™2 Transfer Stacks ........................................................ 57
Additional products ............................................................ 57
Precast gels and premade buffers ................................................ 58
■APPENDIX G Safety ..................................................... 59
Instrument safety .............................................................. 59
General .................................................................. 59
Physical injury ............................................................ 59
Electrical safety ........................................................... 60
Cleaning and decontamination ............................................... 61
Instrument component and accessory disposal ................................ 61
Safety and electromagnetic compatibility (EMC) standards .......................... 61
Symbols on instrument ..................................................... 62
Safety standards ........................................................... 62
EMC ...................................................................... 62
Environmental design standards ............................................. 63
Chemical safety ................................................................ 64
Biological hazard safety ......................................................... 65
■APPENDIX H Documentation and support ............................ 66
Customer and technical support ................................................. 66
Limited product warranty ....................................................... 66
Contents
iBlot
™
2 Dry Blotting System User Guide
5
Product information
Product contents
This manual is supplied with the iBlot™ 2 Gel Transfer Device (Cat. no. IB21001).
The contents of the iBlot™ 2 Gel Transfer Device are listed below. See “iBlot™ 2 Gel
Transfer Device specications“ on page 55 for specications and description of the
iBlot™ 2 Gel Transfer Device.
Component Quantity
iBlot™2 Gel Transfer Device 1
Blotting Roller 1
Stylus 1
Power Cord 1
Power Adapters (North America/Japan,
Europe, UK)
1 set
Before you begin using this product, or any installation or service operation, please
read the following safety information. Aention to these warnings will help prevent
personal injuries and damage to the products.
It is your responsibility to use the product in an appropriate manner. This product is
designed for use solely in laboratory environments, and must not be used in any way
that may cause personal injury or property damage.
You are responsible if the product is used for any intention other than its designated
purpose or in disregard of Thermo Fisher Scientic instructions. Thermo Fisher
Scientic shall assume no responsibility for such use of the product.
The product is used for its designated purpose if it is used in accordance with its
product documentation and within its performance limits.
Using the product requires technical skills and a basic knowledge of English. It is
therefore essential that only skilled and specialized sta or thoroughly trained
personnel with the required skills be allowed to use the product.
Keep the basic safety instructions and the product documentation in a safe place and
pass them on to the subsequent users.
Applicable local or national safety regulations and rules for the prevention of
accidents must be observed in all work performed.
1
Types of products
iBlot™2 Gel
Transfer Device
contents
Before starting
6
iBlot
™
2 Dry Blotting System User Guide
The iBlot™ 2 Gel Transfer Device complies with the TUV Rhineland North America
Inc. safety requirements, part 15 of the FCC rules, and the European Community
Safety requirements. Operation of the iBlot™ 2 Gel Transfer Device is subject to the
conditions described in this manual.
Operation of the iBlot™ 2 Gel Transfer Device is subject to the following conditions:
• Indoor use
• Altitude below 2,000 meters
• Temperature range: 5 to 40°C
• Maximum relative humidity: 80% (maximum relative humidity 80% for
temperatures up to 31°C, decreasing linearly to 50% relative humidity at 40°C)
• Installation categories (over voltage categories) II; Pollution degree 2
• Mains supply voltage uctuations not to exceed 10% of the nominal voltage (100–
240 V, 50/60 Hz, 6.3 A)
• Mains plug is a disconnect device and must be easily accessible.
• Do not aempt to open the iBlot™ 2 Gel Transfer Device. To honor the
warranty,iBlot™ 2 Gel Transfer Device can only be opened and serviced by
Thermo Fisher Scientic.
• The protection provided by the equipment may be impaired if the equipment is
used in a manner not specied by Thermo Fisher Scientic.
• The device must be connected to a mains socket outlet with protective earthing
connections.
• Ventilation requirements: Room ventilation
• Do not use if device becomes cracked or broken in any area.
The product may be installed only under the conditions and in the positions specied
by Thermo Fisher Scientic.
Following are the required operating position and conditions:
• Do not place the product in an area where it will be subject to vibration.
• Do not place the product on surfaces, vehicles, cabinets or tables that for reasons
of weight or stability are unsuitable for this purpose.
• Do not place the product on heat-generating surface or near heat emiing devices
such equipment racks or heaters. Verify that there is sucient clearance between
the product and any other system that may exhaust warm air.
• The product's ventilation should not be obstructed. If proper ventilation is not
provided it can result in electric shock, re and/or serious personal injury or
death.
• The product is for indoor use only.
• Use only with suitably rated mains supply cord (having 3 conductors, min. 16
AWG or 1.5 mm2, min. 300V, Harmonized Type for Europe and UL Listed/CSA
Certied for North America, with molded plug rated min. 10A).
• A tolerance of ±10% shall apply to the nominal input voltage and ±3% to the
nominal frequency, over voltage category 2.
• Maximum operating altitude 2,000 m asl. Maximum transport altitude 4,500 m
asl.
Installing the
instrument
Chapter 1 Product information
Product contents
1
iBlot
™
2 Dry Blotting System User Guide
7
In the event of an equipment malfunction, it is the responsibility of the customer to
report the need for service to Thermo Fisher Scientic or to one of the authorized
agents. For service information, contact Technical Support.
Servicing of this device is to be performed by trained service personnel only.
Unpacking instructions
Examine the unit carefully for any damage incurred during transit. File any damage
claims with the carrier. The warranty does not cover in-transit damage.
• Remove the roller and stylus from the Styrofoam packaging.
• Remove the device from the Styrofoam packaging.
• Remove the protective lm from the touch screen.
iBlot™2 Gel Transfer Device
The front-top view showing various parts of the iBlot™ 2 Gel Transfer Device is shown
below.
Service operation
requirements
Upon receiving the
instrument
Unpacking the
iBlot™2 Gel
Transfer Device
Front view
Chapter 1 Product information
Unpacking instructions
1
8
iBlot
™
2 Dry Blotting System User Guide
A rear view showing various parts of the iBlot™ 2 Gel Transfer Device is shown below.
A side view showing various parts of the iBlot™ 2 Gel Transfer Device is shown below.
About the system
The iBlot™ 2 Dry Bloing System consists of the iBlot™ 2 Gel Transfer Device and
associated iBlot™ 2 Transfer Stacks (sold separately). The iBlot™ 2 Gel Transfer Device
has a unique design, which, in conjunction with the patented gel matrix technology of
the iBlot™ 2 Transfer Stacks, results in a shortened distance between electrodes, high
eld strength, and high currents to reduce transfer times when bloing proteins onto
membranes.
Western bloing of proteins from midi- or mini-sized polyacrylamide gels onto
nitrocellulose or PVDF membranes within 7 minutes can be performed with iBlot™ 2
Transfer Stacks.
See the next page to understand how the iBlot™ 2 Dry Bloing System works and
“Description of parts“ on page 12 for details on various parts of the system.
Rear view
Side view
iBlot™2 Dry
Blotting System
Chapter 1 Product information
About the system
1
iBlot
™
2 Dry Blotting System User Guide
9
• Pre-programmed (iBlot™ 2 Gel Transfer Device) with six Methods for transfer of
proteins from various gel types in 7–8 minutes
• Built-in safety features in the device enhance user safety
• User-friendly iBlot™ 2 Gel Transfer Device design with an integrated power
supply to avoid inconsistencies associated with the use of an external power
supply
• Fast, reliable protein transfer using iBlot™ 2 Transfer Stacks with integrated
nitrocellulose or PVDF transfer membranes for bloing without the need to
prepare buers
• Compatible for use with Bolt™ Bis-Tris PLUS™, NuPAGE™ Bis-Tris and Tris-
Acetate, Tris-Glycine, Tricine (in mini- and midi gel formats), and E-PAGE™ gels
iBlot™2 Gel Transfer Device
The iBlot™ 2 Gel Transfer Device is a self-contained bloing unit with integrated
power supply used for fast, dry bloing of proteins. See “iBlot™ 2 Gel Transfer
Device“ on page 12 for details.
iBlot™2 Transfer Stacks
The iBlot™ 2 Transfer Stacks are disposable stacks that have integrated PVDF or
nitrocellulose transfer membranes to perform dry bloing of proteins. Each iBlot™ 2
Transfer Stack contains a copper electrode and appropriate cathode and anode buers
in the gel matrix to allow fast, reliable transfer of proteins. See “iBlot™ 2 Transfer
Stacks“ on page 15 for details.
The iBlot™ 2 Dry Bloing System is based on the dry bloing concept, utilizing the
unique, patented gel matrix technology developed for E-Gel™ and E-PAGE™ gels for
the iBlot™ 2 Transfer Stacks.
The iBlot™ 2 Transfer Stack consists of a Boom Stack and a Top Stack sandwiching a
pre-run gel and a nitrocellulose (0.2 µm) or PVDF (0.2 µm) membrane. The iBlot™ 2
Transfer Stack is assembled with the bloing membrane on the anode side, and a pre-
run gel on the cathode side.
Features
System
components
System overview
Chapter 1 Product information
About the system
1
10
iBlot
™
2 Dry Blotting System User Guide
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