GE PhastSystem User manual

Phast System
user manual
automated electrophoresis
um 80-1320-15 Edition AI

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Contents
1. Introduction .................................................................. 5
2. Important safety information ......................................... 9
2.1 Connection to the mains supply ............................... 9
2.2 Safety arrangements ............................................... 9
2.3 Safety precautions .................................................10
3. Description of the system ...............................................11
3.1 The separation and control unit ............................. 11
3.2 The development unit ...........................................15
3.3 PhastGel media and chemicals ...............................1
3.4 Using the keyboard .............................................. 22
3.5 The keyboard ......................................................23
4. Installation .................................................................31
4.1 Unpacking .......................................................... 31
4.2 Cable connections ................................................32
4.3 Turning the system on ..........................................33
4.4 Before use ........................................................... 33
5. Operation ...................................................................35
5.1 Programming separation procedures ...................... 35
5.2 Sample application ............................................... 39
5.3 Running IEF media .............................................. 41
5.4 Running electrophoresis media ..............................46
5.5 Programming development procedures ................... 50
5.6 Running a development method ........................... 54
5.7 Cleaning method .................................................57
5. Temperature compensation ................................... 59
6. Evaluation and presentation of data .............................65
6.1 Preservation ......................................................... 65
6.2 Evaluation ........................................................... 67
. Maintenance and trouble shooting ................................ 3
7.1 Separation and control unit ................................... 74
7.2 Development unit ................................................. 76
7.3 Trouble shooting ..................................................7
8. Ordering information and technical data....................... 9
.1 Ordering information ...........................................79
.2 Technical data ..................................................... 1
1. Contents

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Important user information
Reading this entire manual is
recommended for full
understanding of the use of
this product.
The exclamation mark within an equilateral triangle
is intended to alert the user to the presence of
important operating and maintenance instructions
in the literature accompanying the instrument.
The lightning symbol within an equilateral
triangle is intended to alert the user to the
risk of exposure to high voltages.
The earth (ground) terminal symbol is used
to mark a functional earth terminal.
Should you have any comments on this manual, we
will be pleased to receive them at:
GE Healthcare Bio-Sciences AB
S-751 2 Uppsala
Sweden
GE Healthcare Bio-Sciences AB reserves the right to
make changes in the specifications without prior
notice.
Warranty and Liability
GE Healthcare Bio-Sciences AB guarantees that the
product delivered has been thoroughly tested to
ensure that it meets its published specifications. The
warranty included in the conditions of delivery is
valid only if the product has been installed and used
according to the instructions supplied by
GE Healthcare Bio-Sciences AB.
GE Healthcare Bio-Sciences AB shall in no event be liable
for incidental or consequential damages, including
without limitation, lost profits, loss of income, loss of
business opportunities, loss of use and other related
exposures, however caused, arising from the faulty
and incorrect use of the product.
Trade marks
PhastSystem™, PhastTransfer™ and PhastGel®
are the exclusive trade marks of
GE Healthcare Bio-Sciences AB.
In view of the risk of trade mark
degeneration, it is respectfully suggested that
authors wishing to use these designations refer to
their trade mark status at least once in each article.
Copyright© 1995 GE Healthcare Bio-Sciences AB
All rights reserved. No part of this product may be
reproduced, stored in a retrieval system or transmit-
ted in any form by any means, without permission in
written form from the company.

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1. Introduction
PhastSystem consists of a separation and control unit, a development
unit, high-performance PhastGel® separation media, accessories, and a
technical support package. These components work together to form
a system for fast, high-resolution, and reproducible electrophoresis.
With PhastSystem, isoelectric focusing is as easy to perform as gel
electrophoresis; Coomassie staining is as easy as silver staining. The
schematic diagram below illustrates the steps involved in producing a
finished electrophoresis gel using PhastSystem with PhastGel
separation media.
Flow diagram or PhastSystem
™
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▼
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▼
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▼
Separation and control unit Development unit
Place 1 or 2 gels on Place gel(s) in
the separation bed 3 min. development chamber 1 min.
Load PhastGel Select a programmed
sample applicator(s) 3 min. development method
and press the start button
Select a programmed when method stops 3 -9 min
separation method
and press the start
button
remove the gel(s) and
analyze the results
when alarm sounds 2 -45 min. Total time 32-92 min.
remove the gel(s)
Total time 26-5 min.
or
Specific detection via
conventional methods
e.g. zymograms, auto-
radiography, blotting
The time intervals listed above will depend on the technique that is run.
▼
▼
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1. Introduction

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This users guide includes the following chapters:
Chapter 2: Important safety information.
Chapter 3: Description of the system; introduces you to
PhastSystem.
Chapter 4: Installation; tells you how to install PhastSystem.
Chapter 5: Using the keyboard; prepares you for programming
and running methods.
Separation procedures, shows you how to program
and run separation methods.
Development procedures, shows you how to
program and run development methods.
Chapter 6: Evaluation and presentation of data; gives advice
on drying, mounting, and photographing gels and
describes procedures for molecular weight and
isoelectric point measurement using calibration
proteins.
Chapter 7: Maintenance and trouble shooting; shows you
how to replace and clean certain parts of the
instruments and how to calibrate the temperature
sensors. Provides current trouble shooting
recommendations. If you have any problems
during programming or operation you find all help
messages listed here.
Chapter : Ordering information and technical data; gives you
all information needed to order the products
mentioned in this manual. You will also find a list
of the most common spare parts required for
maintenance of PhastSystem.
A list of the technical data on PhastSystem
instruments and PhastGel media and accessories is
also included.
Chapter 9: Separation technique files; you will find optimized
methods for a number of separation techniques.
Development technique files; you will find
optimized methods for a number of development
techniques.
Application notes; here you can file application and
technical notes covering specific techniques or
application areas.
A technological extension of PhastSystem is PhastTransferTM.
PhastTrans er
PhastTransfer brings speed, reproducibility and convenience to semi-
dry electrophoretic transfer of proteins from PhastGel separation
media to immobilizing membranes. The small format of the gels
together with semi-dry transfer method minimize the amount of
reagents needed for detection. Elution efficiency is greater than 90%
for most protein systems. At 1.0 mA/cm2, high transfer recovery is
obtained, usually within 10–30 minutes.
1. Introduction

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Phastsystem
Fig.1. PhastSystem consists of a separation and control unit, a development unit,
PhastGel separation media, accessories and a technical support package.
1. Introduction

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2. Important safety
information
Voltage selector setting
The instruments are available in two versions: one for 220-230/240
V AC, referred to here as the 220 V model, and one for 100/120 V
AC, referred to here as the 120 V model.
As a safety precaution, check the code number and voltage printed
on the backpanels to ensure you have the correct model for your
local electricity supply.
Code number 1 -101 -23:
Separation and control unit 120 V model
Development unit 120 V model
Code number 1 -101 -24:
Separation and control unit 220 V model
Development unit 220 V model
Set the voltage selectors on the rear panels of the separation and
control unit and development unit according to your local electricity
supply. To do this:
• Check the voltage range of the mains electricity supply.
• Set the voltage selector to the appropriate setting according to
the table below.
Voltage range Voltage selector setting
For12 V model instruments:
9 -11 1
1 8-132 12
For 22 V model instruments:
198-242 22 -23
216-264 24
Important! Always disconnect the mains power cords when
servicing the system.
The operator is protected against high voltage by the separation
compartment lid when an electrophoresis is in progress.
If the lid is opened during a run, the high voltage supply switches off
automatically to eliminate electrical hazard. An alarm will sound
until the lid is closed or until the run is paused.
2.1 Connection to
the mains supply
2.2 Sa ety
arrangements
2. Important safety information

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The voltage supplied by PhastSystem is capable of delivering a lethal
electric shock. The numerous safety devices and circuits built into the
instrument prevent this. The “pause” and “start/stop” keys can also
be pressed to halt the supply of power at any stage of the experiment
or operation of PhastSystem. Nevertheless, in keeping with good
laboratory practise, we advice you to take the following precautions
when dealing with the instrument.
1. Regularly check all insulation cables, take care not to damage
the units, especially the separation compartments lid.
Note: For full safety it is important that the lid is not
tampered with.
2. Ensure that the mains cables are plugged into fully grounded
mains outlets.
3. Allow only authorized service representatives to service or work
on the electrical circuitry of PhastSystem.
4. Avoid spilling buffers or other conduction liquids onto the
instrument.
5. Allow the ventilation slots (situated at the rear of PhastSystem)
to have free access to a good flow of air.
2.3 Sa ety
precautions
2. Important safety information