Datasheet k systému Ettan IPGphor II (EN)

Datasheet k systému Ettan IPGphor II (EN)
IPGphor_Brochure.qxd 6/7/2003 16:39 Page 21
First-Dimension Isoelectric Focusing System
JASON & JASON VISUAL COMMUNICATIONS
Ettan
IPGphor II
WORK
designed to
18-1176-09 AA
IPGphor_Brochure.qxd 6/7/2003 16:39 Page 22
Amersham Biosciences continues to set the standard
for the highest performance in 2-D electrophoresis
with an ever-increasing product portfolio that saves
time, improves resolution, and increases capacity and
throughput—while always focusing on achieving
the highest reproducibility and the highest quality
results. Ettan™ IPGphor™ II Isoelectric Focusing
Unit and Ettan IPGphor Manifold are the latest in
the company’s state-of-the-art 2-D electrophoresis
platform that have been designed to work in today’s
most demanding proteomics applications.
IPGphor_Brochure.qxd 6/7/2003 16:39 Page 1
Ettan IPGphor II
A professional solution for professionals
Ettan IPGphor II overcomes the most
common hurdles of existing technology
to consistently deliver high-throughput,
speed, reproducibility, and high proteinloading capacity.
The original IPGphor system, designed
for immobilized pH gradient IEF, has
been upgraded and offers significant
improvements. Ettan IPGphor II is
easier to use, capable of generating
fast, reproducible runs, and now features PC control.
Ettan IPGphor II Control Software enables
creation, storage, and editing of protocols.
The new Ettan IPGphor Manifold is a
high-throughput accessory that is
optimized for first-dimension isoelectric
focusing of micropreparative loads of
proteins with speed, convenience, and
reproducibility.
The extensive range of Immobiline™
DryStrip gels is now bar coded to enable
sample and lot tracking.
1
IPGphor_Brochure.qxd 6/7/2003 16:39 Page 2
New design
performance
for improved
The newly designed Ettan IPGphor II
is a fully integrated isoelectric focusing
system optimized to deliver ease-ofuse, reproducibility, and efficient
operation. The large graphical display
accommodates multiple (up to four)
lines of text for fast and easy programming. Up to10 protocols (nine
steps each) can be saved, retrieved
and easily edited on the instrument.
Any number of protocols can be
stored on a connected PC running
IPGphor II Control Software and
uploaded to the instrument instantly.
Increased safety features ensure safe
high-voltage runs, and with total
start/stop control from your PC, IEF
runs have never been easier.
• Integral 8000 V power supply
• Peltier solid-state temperature
control (18–25 °C)
• Accomodates one manifold tray or
1-12 stripholders for 7, 11, 13,18
and 24 cm IPG strips
Easy access to
serial port
Enhanced user interface
with four-line display
Leveling feet
• Programmable controller for
voltage, temperature, and time
IPGphor_Brochure.qxd 6/7/2003 16:39 Page 3
Enhanced software for improved control
Ettan IPGphor II Control Software can be used to control up to four Ettan
IPGphor II units simultaneously, each running a different set of run parameters.
• Create, save and edit protocols
• Monitor voltage, current,
and volt-hours of the run and
generate graphical display as
the run proceeds
• Open and view stored log
files of previous runs
• Start, stop, and pause the
Ettan IPGphor II
• Generate status report on
Ettan IPGphor II (instantaneous
run condition report on request)
• Enable Web browser remote
monitoring of PC control unit
• Export log files to programs
such as Microsoft® Excel®.
B
C
A
A – Trend lines B – Scales C – Voltage, current, and volt-hours
• Create professional reports
that can be saved, printed,
and exported
2
3
manifold improved strip processing
IPGphor_Brochure.qxd 6/7/2003 16:39 Page 4
The Ettan IPGphor Manifold is a high-throughput cup-loading accessory
for first-dimension isoelectric focusing of proteins on IPG strips. The
newly designed manifold meets the ever-growing demands of proteomics
research and delivers high-resolution micropreparative protein analysis. Ettan
IPGphor Manifold simplifies isoelectric focusing of multiple samples and reduces
first-dimension hands-on time to a minimum. This significantly speeds up the
processing of micropreparative loads in first dimension analysis.
The unique design of the manifold handles isoelectric focusing and subsequent
equilibration for up to 12 IPG strips. Designed for maximum flexibility, the
manifold can accommodate all IPG strip sizes of 7, 11, 13, 18, and 24 cm. The
manifold’s versatility makes it possible to use for equilibrating the IPG strips after
first-dimension IEF, which saves substantial time and reduces the number of
parts. Ettan IPGphor Manifold comes with a complete set of accessories to run
10 full runs of 12 strips each, including forceps, cleaning brush, IPGphor
zcleaning solution and a spirit level. The manifold is also compatible with the
first-generation Ettan IPGphor.
IPGphor_Brochure.qxd 6/7/2003 16:39 Page 5
Designed for your advantage
Unique cup-loading application
Electrode assembly
The unique cup-based sample application improves
protein-focusing patterns, particularly in basic IPG strips,
and accommodates both anodic and cathodic loading.
Under conditions where substantial water transport, or
electroendosmosis, accompanies focusing, such as with
protein loads in excess of 1 mg, the face-up mode
frequently yields improved resolution.
State-of-the-art electrode and sample cup design
ensure consistent quality and superior results for
micropreparative runs. The movable electrodes can be
easily adjusted to any strip length (from 7–24 cm), and
can be placed on any of the assigned positions along the
manifold where the electrode pins will make electrical
contact with the power supply on the bed of IPGphor.
Special hold-down teeth apply pressure on the paper
bridges when the manifold is used to run large protein
loads by paper bridge loading the samples, and specially
designed cams on the sides of the electrodes secure the
electrodes in place.
Manifold tray for easy placement
The manifold tray base is made of a thermally conductive
aluminum oxide ceramic that rapidly dissipates heat
to avoid “hot spots.” A further special coating of the surface eliminates protein absorption. The manifold tray
allows simple and accurate placement of IPG strips, with
protrusions along the numbered inner channels that keep
IPG strips straight and centered.
Sample cups
Sample cups can be used to load more protein in discrete
sample zones and can run the IPG strip “gel-side-up.” Each
cup can hold sample volumes of up to 150 µl, and can be
properly positioned and sealed over the IPG strips with an
innovative sample cup insertion tool.
Electrode wicks and paper bridges
Pre-cut electrode pads and paper bridges are convenient
and save valuable time. The wicks absorb excess water,
salts, and proteins while the paper bridges can be used to
load large sample volumes, as well as ensure that the wicks
are not contaminated with human proteins during cutting.
Ettan IPGphor II Strip Holders
The innovative IPGphor Strip Holder serves as both a
rehydration and focusing chamber for individual IPG
strips. When the sample is included in the rehydration
solution, it is loaded into the gel by absorption during
the rehydration. Since the rehydrated gel is in direct
contact with electrodes built into the strip holder, it is
placed in position to run without further handling.
The base of the strip holder is made from the same
thermally conductive aluminum oxide ceramic as the
manifold and has platinum electrodes at each end.
The transparent strip holder cover enables easy visual
monitoring of rehydration and focusing progress.
4
5
IPGphor_Brochure.qxd 6/7/2003 16:39 Page 6
Immobiline DryStrip
with IPG Buffer
For unmatched reproducibility, resolution, and accuracy
• High batch-to-batch
reproducibility
• Superior accuracy
For excellent first-dimension separation, the best choice is Immobiline DryStrip
gels with IPG Buffer. The IPG strip delivers unrivaled resolution and high reproducibility, without compromising throughput, loading capacity, or maximum
running conditions. The IPG Buffers ensure stable pH gradients to deliver the
• Optimized running
conditions
• Excellent resolution with
no background staining
highest degree of accuracy, and the buffers eliminate the problem of background staining. The high-quality first-dimension results from the Ettan IPGphor
II platform ensures that spots in the second dimension are well separated.
And the growing range of precast 24 cm IPG strip gels with broad, medium, and
narrow pH ranges simplifies and accelerates development of concise targeted
• High sample loading
strategies for analyzing complex cell extracts.
Shorter strips for rapid
screening, or when only
the most abundant
proteins are of interest.
Longer strips for
maximal resolution
and loading capacity.
To overview total
protein distribution,
use pH interval of
3–10.
IPGphor_Brochure.qxd 6/7/2003 16:39 Page 7
Barcode for IPG strips and DALT Gel 12.5
• Convenient sample and lot tracking
• Minimizes risk of mixing samples up
• Easy to transfer with a barcode reader to
electronic laboratory journal pages composed
in popular office programs, such as Microsoft
WordPad, Microsoft Word, Microsoft Excel
• Supports sample tracking in Ettan 2D-MS
LWS software
Flexibility for the results
you need
For increased resolution between
pH 5 and 7, use a nonlinear gradient,
pH 3–10 strip to distribute the
proteins more evenly over the gel
Combine pH 4–7 and pH 6–11 (or
pH 4–7 and pH 6–9) for more detail
Use narrow pH range (1 pH unit) IPG
strips to closely study proteins in the
regions of interest
IPG pH range
Sample loaded
3–10
40 µg
5.0–6.0*
120 µg
4–7
80 µg
*Using pH 5.0–6.0 reveals more spots at greater distances apart than when using pH 3–10
24 cm
3.5–4.5
4.0–5.0
4.5–5.5
5.0–6.0
5.5–6.7
3–7 NL
4–7
6–9
3–10 NL
3–10
18 cm
3.5–4.5
4.0–5.0
4.5–5.5
5.0–6.0
5.5–6.7
4–7
6–9
6–11
3–10 NL
3–10
13 cm
4–7
6–11
3–10 NL
3–10
11 cm
4–7
6–11
3–10
17 cm
4–7
6–11
3–10 NL
3–10
8
9
10
11
12
3–10 NL
7
6–11
6
IPG Buffer
4–7
5
5.0–6.0
4
4.5–5.5
3
3.5–5.0
2
pH range
5.5–6.7
Immobiline DryStrip
pH range
Resolution and
loading capacity
6
7
IPGphor_Brochure.qxd 6/7/2003 16:39 Page 8
step 1
step 2
step 3
High-throughput
micropreparative
runs made easy
First-dimension isoelectric focusing in micropreparative
scale of multiple samples can be challenging. But with the
improved Ettan IPGphor II, with its innovatively designed
Manifold, first-dimension separation with high resolution is
easy. With the gel side facing up, samples are applied in a
localized region through an open-bottom loading cup. This
cup loading technique improves protein-focusing patterns,
particularly on basic IPG strips (pH 6–9 and 6–11).
IPGphor_Brochure.qxd 6/7/2003 16:39 Page 9
step 4
step 5
step 6
step 7
step 8
Step 1 – Rehydrate IPG strips
Step 6 – Position Electrode Assembly
Rehydrate Immobiline DryStrip gels, with the gel side
down, in the appropriate volume of rehydration solution,
using Immobiline DryStrip Reswelling Tray.
Allow the IPG strips to rehydrate overnight (10–20 h).
Slide an electrode assembly over the top of all the pads.
Swivel the cams into the position under the external lip
of the Manifold to seat the electrode in place.
Step 2 – Position Manifold
Load the samples into the sample cups, up to a maximum of 150 µl. Check to make sure that the samples are
completely covered with strip cover fluid.
Position the Manifold on Ettan IPGphor II. The small
T-shaped protrusion fits into the cutout section of the
Ettan IPGphor II bed making positioning easy.
Step 3 – Transfer to Cup Loading Manifold
Pour the appropriate volume of Immobiline DryStrip
Cover Fluid evenly in all the channels. Transfer the IPG
strips to the Ettan IPGphor II Cup Loading Manifold.
Place them face up in the tray with the anodic (+)
end of the strip resting on the appropriate mark etched
on the bottom of the Manifold track. Fill the Manifold
with strip cover fluid. The Manifold can be used for
cathodic loading as well.
Step 7 – Load and Cover Samples
Step 8 – Set Program Parameters and Run
Close the Ettan IPGphor II cover. Select program and
desired run parameters and begin the run.
Post Electrophoresis – Equilibration
At the end of a run, the Manifold may be used to
equilibrate the strips to prepare for second-dimension
electrophoretic analysis.
1
Step 4 – Seat Cups in Track
Place a strip of cups in the appropriate position. The
convenient seating tool enables you to push the cups
down so that they are properly seated at the bottom
of the track.
Step 5 – Moisten and Place Electrode Pads
Wet the pre-cut electrode pads with deionized water and
blot until they are almost completely dry. Place the pads
on the IPG strips.
2
3
Remove the electrode assembly, wicks and bridges
from the anodic (+) side of the strips and discard the
strip cover fluid.
Add equilibration solution (5–6 ml) to each track and
incubate the IPG strips in the solution.
Discard the equilibration solution and either add
other solutions as needed, or place strips on top of
second-dimension gels.
8
9
IPGphor_Brochure.qxd 6/7/2003 16:39 Page 10
1 Preparative run: Immobiline
DryStrip pH 3-10, 24cm. Sample:
1.7 mg E. coli containing 50 mM Tris.
Samples were rehydrated with
DeStreak™ Rehydration Solution, transferred to the Manifold and run on the
Ettan IPGphor II for first-dimension IEF.
Sample was focused for 124 kVh and
run on Ettan DALTtwelve Large
Vertical Electrophoresis System for
the second dimension.
1
Enhanced
2-D separations
2 Preparative run: Immobiline
DryStrip pH 4.5–5.5, 24 cm, rehydrated in DeStreak Rehydration Solution.
Sample: 85 µl mouse liver extract
containing 2.5 mg proteins diluted
with 300 µl DeStreak Rehydration
Solution and applied with a paper
bridge at the anode. Total run time
was 95 kVh. Coomassie™ stained.
2
3
Preparative run: Immobiline
DryStrip pH 4.5–5.5, 24 cm.
Sample 85 µl mouse liver extract
containing 2.5 mg protein. Sample
applied by rehydration loading 20
mM DTT in sample and rehydration
solution. Total run time was 95 kVh.
Coomassie stained.
4
3
4
Analytical run: Anodic cup application.
Immobiline DryStrip pH 6–11, 18 cm rehydrated in DeStreak Rehydration Solution.
Sample: 60 µl mouse liver extract
containing 75 µg proteins. Total run
time 26 kVh. Silver stained.
IPGphor_Brochure.qxd 6/7/2003 16:39 Page 11
for better protein identification
5 Preparative rehydration loading:
Immobiline DryStrip pH 5.0–6.0, 24
cm, DeStreak Rehydration Solution.
Sample: 2 mg E. coli proteins applied in
the rehydration solution. Total run time
95 kVh. Coomassie stained.
6 Preparative run, paper bridge
sample application: Immobiline
DryStrip pH 5.0–6.0, 24 cm,
DeStreak Rehydration Solution.
Sample: 2.4 mg mouse liver proteins
diluted in 320 µl DeStreak Rehydration
solution applied at the anode. Total
run time 95 kVh. Coomassie stained.
7 Preparative run, paper-bridge
sample application: Immobiline
DryStrip pH 6–9, 24 cm, DeStreak
rehydration solution.
Sample: 1.4 mg mouse liver proteins in
270 µl DeStreak Rehydration Solution
applied at the anode. Total run time
95 kVh. Coomassie stained.
5
6
7
10
11
IPGphor_Brochure.qxd 6/7/2003 16:39 Page 12
Technical Information
Ettan IPGphor II
Working surface
Electrode areas
Gold-plated copper
Operating environment
Temperature
15–35 °C
Strip Holder capacity
Up to 12 fixed-length,
on one manifold tray
Relative humidity
0–70%
Platform temperature
15–30 °C
Power requirements
Line voltage
115/230 V~
Frequency
50/60 Hz
Power consumption
100 W
Strip holders
Material
Aluminum oxide
ceramic base,
acrylic cover,
polycarbonate,
platinum on titanium
Power outputs
Voltage
0–8000 V,
resolution: 10 V
Holders
For IPG strip lengths:
7, 11, 13, 18, and 24 cm
Current
Cup loading manifold
For all strip lengths, cup
and bridge loading
0–1.5 mA,
resolution: 1 µA
Power
12 W max.
Seven-key
membrane keypad
Physical parameters
Dimensions (h x w x d)
14 x 25 x 46 cm
Liquid crystal display (LCD) Four lines, 24
characters per line
Weight
6.8 kg (15 lbs)
Programmable parameters
Safety
Safety features automatic voltage cutoff
when safety lid is opened
User interface
Control panel
Rehydration time,
platform temperature,
maximum current limit
per strip, step voltage,
step voltage change
pattern, and step
duration
Protocol capacity
10, with up to
9 steps each
Serial port
RS232C, 1200 or 9600
baud, 8 data bits,
1 stop bit, no parity,
no flow control
Safety certifications CE, UL, CSA
Ettan IPGphor II Control Software
Software requirements
Hardware requirements
PC Windows® 98 SE (Second Edition) or higher, Windows NT® or Windows 2000.
Serial port RS232. If multiple Ettan IPGphor II units are to be controlled,
an RS232 expansion box is required.
IPGphor_Brochure.qxd 6/7/2003 16:39 Page 13
new!
How to
order
The Ettan IPGphor II platform is
available in three application-based options
for High-Throughput Analytical and Micropreparative
Protein Analysis. Complete solution.
•
•
•
•
•
80-6505-03 Ettan IPGphor II Isoelectric Focusing Unit
80-6498-38 Ettan IPGphor Manifold - complete**
80-6465-32 Reswelling Tray, 7–24 cm
Range of ceramic Strip Holders to run 7,11,13,18 or 24 cm IPG strips***
Range of IPG strips and buffers*
for High-throughput Micropreparative Applications
•
•
•
•
80-6505-03 Ettan IPGphor II Isoelectric Focusing Unit
80-6498-38 Ettan IPGphor Manifold - complete**
80-6465-32 Reswelling Tray, 7–24 cm
Range of IPG strips and buffers*
for Analytical Study of Protein Profiles
• 80-6505-03 Ettan IPGphor II Isoelectric Focusing Unit
• Range of ceramic Strip Holders to run 7,11,13,18 or 24 cm IPG strips***
• Range of IPG strips and buffers*
* see Web site for more details
** includes accessories for 10 complete runs of 12 IPG strips
*** essential accessories (see below)
essential accessories
Strip Holders
Product
7 cm,
11 cm,
13 cm,
18 cm,
24 cm,
7 cm,
11 cm,
13 cm,
18 cm,
24 cm,
complete
complete
complete
complete
complete
complete
complete
complete
complete
complete
Other Essentials
Quantity
6
6
6
6
6
1
1
1
1
1
Code Number
80-6416-11
80-6416-30
80-6416-49
80-6416-68
80-6469-88
80-6416-87
80-6417-06
80-6417-25
80-6417-44
80-6470-07
Strip Holder Covers
7
11
13
18
24
cm
cm
cm
cm
cm
2
2
2
2
2
80-6455-44
80-6455-63
80-6455-82
80-6456-01
80-6479-76
Product
Quantity
Immobiline DryStrip
Reswelling Tray, 7–18 cm
1
Immobiline DryStrip
Reswelling Tray, 7–24 cm
1
Strip Holder Cleaning
Solution 950 ml
1
2-D Electrophoresis
Principles and Methods
1
Equilibration tube set
for 24 cm IPG Strips
12
IEF electrode strips
100
Manifold sample cups (6)
20
Pre-cut electrode wicks (6)
40
Pre-cut paperbridge pads (6)
20
Manifold electrode set
1
Lid adapter
1
Cleaning brush
1
Spirit level
1
Forceps SS
1
Code Number
80-6371-84
80-6465-32
80-6452-78
80-6429-60
80-6467-79
18-1004-40
80-6498-95
80-6499-14
80-6499-33
80-6498-76
80-6499-71
80-6505-98
80-6194-19
80-6506-17
IPGphor_Brochure.qxd 6/7/2003 16:39 Page 20
For more information about our IPGphor platform, visit:
www.designed-to-work.com
Denmark
Tel: 4516 2400
Fax: 4516 2424
Latin America
Tel: +55 11 3933 7300
Fax: +55 11 3933 7306
South East Asia
Tel: 60 3 8024 2080
Fax: 60 3 8024 2090
Australia
Tel: +61 2 9899 0999
Fax: +61 2 9899 7511
Finland & Baltic Nations
Tel: +358 (0)9 512 39 40
Fax: +358 (0)9 512 17 10
Middle East and Africa
Tel: +30 210 96 00 687
Fax: +30 210 96 00 693
Spain
Tel: 93 594 49 50
Fax: 93 594 49 55
Austria
Tel: 01 576 0616 19
Fax: 01 576 0616 27
France
Tel: 01 69 35 67 00
Fax: 01 69 41 96 77
Netherlands
Tel: 0165 580 410
Fax: 0165 580 401
Sweden
Tel: 018 612 19 00
Fax: 018 612 19 10
Belgium
Tel: 0800 73888
Fax: 03 272 1637
Germany
Tel: 0761 4903 490
Fax: 0761 4903 405
Norway
Tel: 23 18 58 00
Fax: 23 18 68 00
Switzerland
Tel: 0848 802 28 12
Fax: 0848 802 28 13
Canada
Tel: 1 800 463 5800
Fax: 1 800 567 1008
Italy
Tel: 02 27322 1
Fax: 02 27302 212
Portugal
Tel: 21 417 70 35
Fax: 21 417 31 84
UK
Tel: 0800 616 928
Fax: 0800 616 927
Central, East &
South East Europe
Tel: +43 1 982 3826
Fax: +43 1 985 8327
Japan
Tel: 81 3 5331 9336
Fax: 81 3 5331 9370
Russia & other C.I.S.&N.I.S.
Tel: +7 (095)232 0250
Fax: +7 (095)230 6377
USA
Tel: 800 526 3593
Fax: 877 295 8102
Ettan, DeStreak, Immobiline, and IPGphor are trademarks of Amersham Biosciences Limited. Amersham and Amersham Biosciences are trademarks of
Amersham plc. © Amersham Biosciences UK Limited 2003 – All rights reserved. Coomassie is a trademark of ICI plc. Excel, Microsoft, NT, and Windows are
trademarks of Microsoft Corporation. All goods and services are sold subject to the terms and conditions of sale of the company within the Amersham
Biosciences group which supplies them. A copy of these terms and conditions is available on request. Amersham Biosciences AB SE-751 84 Uppsala, Sweden.
Amersham Biosciences UK Limited Amersham Place Little Chalfont Buckinghamshire, England HP7 9NA. Amersham Biosciences Corp. 800 Centennial Avenue
PO Box 1327 Piscataway, NJ 08855 USA. Amersham Biosciences Europe GmbH Munzinger Strasse 9 D-79111 Freiburg, Germany. Amersham Biosciences K.K.
Sanken Building, 3-25-1, Hyakunincho, Shinjuku-ku, Tokyo.
JASON & JASON VISUAL COMMUNICATIONS
Asia Pacific
Tel: +852 2811 8693
Fax: +852 2811 5251
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