cytiva Ettan IPGphor Cup Loading Manifold User manual

Ettan IPGphor Cup Loading
Manifold
User Manual
cytiva.com

Table of Contents
1Introduction ........................................................................................................ 3
1.1 Important user information ....................................................................................................................... 4
2 Description .......................................................................................................... 6
2.1 Overview ............................................................................................................................................................ 7
2.2 Accessories ...................................................................................................................................................... 12
3 Operation ............................................................................................................ 13
3.1 Instructions ...................................................................................................................................................... 14
3.2 Run conditions ................................................................................................................................................ 21
4 Maintenance ....................................................................................................... 22
5 Troubleshooting ................................................................................................. 24
6 Reference information ....................................................................................... 27
Table of Contents
2 Ettan IPGphor User Manual 11003460 AE

1 Introduction
Purpose of the User Manual
The User Manual provides you with the instructions needed to use Ettan™ IPGphor™
Cup Loading Manifold together with an Ettan IPGphor unit.
In this chapter
Section See page
1.1 Important user information 4
1 Introduction
Ettan IPGphor User Manual 11003460 AE 3

1.1 Important user information
Read this before operating Ettan
IPGphor unit
All users must read the entire Ettan IPGphor Operating Instructions before
installing, operating or maintaining the instrument. Always keep the Operating
Instructions at hand when using Ettan IPGphor.
Do not operate Ettan IPGphor in any other way than described in the user documenta-
tion. If you do, you may be exposed to hazards that can lead to personal injury and you
may cause damage to the equipment.
Intended use
The Ettan IPGphor Cup Loading Manifold in combination with Cytiva's Ettan IPGphor,
Ettan IPGphor II and Ettan IPGphor 3 Isoelectric Focusing Units, is suitable for first-
dimension isoelectric focusing (IEF) of proteins on Immobiline™ DryStrip immobilized
pH gradient (IPG) gel strips 3 mm wide, from 7 cm to 24 cm long.
Ettan IPGphor is intended to be used as a laboratory device for research purposes.
Ettan IPGphor shall not be used in any clinical procedures, or for diagnostic purposes.
Definitions
This user documentation contains safety notices (WARNING, CAUTION, and NOTICE)
concerning the safe use of the product. See definitions below.
WARNING
WARNING indicates a hazardous situation which, if not avoided,
could result in death or serious injury. It is important not to proceed
until all stated conditions are met and clearly understood.
CAUTION
CAUTION indicates a hazardous situation which, if not avoided,
could result in minor or moderate injury. It is important not to
proceed until all stated conditions are met and clearly understood.
1 Introduction
1.1 Important user information
4 Ettan IPGphor User Manual 11003460 AE

NOTICE
NOTICE indicates instructions that must be followed to avoid
damage to the product or other equipment.
Notes and tips
Note: A note is used to indicate information that is important for trouble-free and
optimal use of the product.
Tip: A tip contains useful information that can improve or optimize your proce-
dures.
Typographical conventions
Software items are identified in the text by bold italic text.
Hardware items are identified in the text by bold text.
In electronic format, references in italics are clickable hyperlinks.
1 Introduction
1.1 Important user information
Ettan IPGphor User Manual 11003460 AE 5

2 Description
About this chapter
This section provides an overview of the Ettan IPGphor Cup Loading Manifold unit.
In this chapter
Section See page
2.1 Overview 7
2.2 Accessories 12
2 Description
6 Ettan IPGphor User Manual 11003460 AE

2.1 Overview
Introduction
The Ettan IPGphor Cup Loading Manifold, in combination with Cytiva's Ettan IPGphor,
Ettan IPGphor II and Ettan IPGphor 3 Isoelectric Focusing Units, is suitable for first-
dimension isoelectric focusing (IEF) of proteins on Immobiline DryStrip immobilized pH
gradient (IPG) gel strips 3 mm wide, from 7 cm to 24 cm long. IEF is performed with the
gel side facing up.
Samples up to 150 μl are applied in an open-bottom loading cup. Cup loading has been
found to improve protein focusing patterns, particularly on basic IPG strips (pH 6–9 and
6–11). The manifold can accommodate anodic and cathodic cup loading.
Larger volumes of the sample, 375-500 μl, are applied with paper bridge application.
Sample volumes up to the volume of the strip (18 cm strip = 340 μl) can be applied by in
gel rehydration loading.
Illustration of the Ettan IPGphor Cup
Loading Manifold
The illustration below shows the main components of the Ettan IPGphor Cup Loading
Manifold:
1
2
3
5
6
8
9
4
7
10
2 Description
2.1 Overview
Ettan IPGphor User Manual 11003460 AE 7

Part Function Part Function
1 Manifold 6 Pre-cut paper bridges
2 Pre-cut electrode wicks 7 Sample cup insertion tool
3 Electrode assembly 8 Cleaning solution
4 Sample cups 9 Forceps
5 Cleaning brush 10 Spirit level
Manifold
Ceramic Manifold
The manifold (see following figure) is made of aluminum oxide ceramic for efficient
heat transfer and temperature control during IEF. Protrusions along the channel inside
the manifold align the rehydrated IPG strip, keeping it straight and centered when
placed inside the manifold. The manifold has a coating to minimize protein adsorption.
Because some cleaning agents can remove this coating, clean the manifold only with
the Ettan IPGphor Strip Holder Cleaner.
Note: The manifold is fragile and should be handled with care.
Manifold Light
The manifold light is made of the polymers PC (polycarbonate) and ABS (acrylonitrile-
butadiene-styrene).
The manifold light's flat design is important for the heat transfer. If exposed to very
high temperatures, the flatness could be affected. Refer to Manifold Light, on page 23
regarding cleaning temperature.
General information
IPG strips from 7 cm to 24 cm long may be used. The unit may also be used for IPG strip
equilibration prior to second dimension electrophoresis. Forcep wells are located at
the ends of the channels to aid strip removal after electrophoresis.
2 Description
2.1 Overview
8 Ettan IPGphor User Manual 11003460 AE

forcep wells
channel
numbering centering
protrusion
Electrode assembly
The movable electrodes, as shown below, can be placed along the manifold to fit the
various length of Immobiline Dry Strip. Electrode pins make electrical contact with the
power supply pads on the bed of the Ettan IPGphor. The electrodes have a platinum
wire that is strung across the bottom of the electrode teeth to provide electrical
contact to the IPG strips. The hold down teeth are found along the opposite length of
the electrode. They are used to apply pressure on the paper bridges when the manifold
is used for bridge loading. Cams on the sides of the electrode secure the electrodes in
place and must be in the closed position during IEF.
cam in open
position
hold down
teeth
electrode
teeth
cam in closed
position
2 Description
2.1 Overview
Ettan IPGphor User Manual 11003460 AE 9

Sample cups
Sample cups, as shown in the figure below, can be placed along the length of the Ettan
IPGphor manifold that is not blocked by a centering protrusion. The sample cups are
supplied in strips of 6 for easy handling and placement. If fewer than 6 cups are
required, they may be easily separated by cutting the thin plastic bridging the strip of
cups together. The sample cups can accommodate sample volumes of up to 150 μl. For
proper sealing of the cup to the gel, all of the feet of the sample cup must rest on the
bottom of the channel and all cup arms must be fully pressed down into the channel.
cup feet
cup arms
Sample cup insertion tool
The sample cup insertion tool, shown below, is supplied with the manifold kit and refill
packages of sample cups. It is used for proper placement and sealing of the cup over
the IPG strips.
2 Description
2.1 Overview
10 Ettan IPGphor User Manual 11003460 AE