cytiva Ettan IPGphor Cup Loading Manifold User Manual
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cytiva Ettan IPGphor Cup Loading Manifold User Manual

cytiva Ettan IPGphor Cup Loading Manifold User Manual

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Ettan IPGphor Cup Loading
Manifold
User Manual
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Summary of Contents for cytiva Ettan IPGphor Cup Loading Manifold

  • Page 1 Ettan IPGphor Cup Loading Manifold User Manual cytiva.com...

  • Page 2: Table Of Contents

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

  • Page 3: Introduction

    1 Introduction 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 Important user information Ettan IPGphor User Manual 11003460 AE...

  • Page 4: Important User Information

    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.
  • Page 5 1 Introduction 1.1 Important user information 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.

  • Page 6: Description

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

  • Page 7: Overview

    2.1 Overview 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.
  • Page 8 2 Description 2.1 Overview Part Function Part Function Manifold Pre-cut paper bridges Pre-cut electrode wicks Sample cup insertion tool Electrode assembly Cleaning solution Sample cups Forceps Cleaning brush 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.
  • Page 9 2 Description 2.1 Overview forcep wells channel centering numbering 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.
  • Page 10 2 Description 2.1 Overview 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.
  • Page 11 2 Description 2.1 Overview IEF electrode paper wicks The small, precut rectangular wicks, shown below, must be placed at both the anodal and cathodal ends of the rehydrated IPG strips just under the electrodes. The wicks absorb excess water, salts, and proteins with pIs that lie outside the pH range of the IPG strip.

  • Page 12: Accessories

    2 Description 2.2 Accessories Accessories Introduction The following accessory items are not supplied with the unit and need to be ordered separately. Immobiline DryStrip Cover Fluid Immobiline DryStrip Cover Fluid is required to ensure that the rehydrated IPG strip gels do not dry out during electrophoresis.

  • Page 13: Operation

    3 Operation Operation About this chapter This section provides instructions on how to operate the Ettan IPGphor Cup Loading Manifold unit. In this chapter Section See page Instructions Run conditions Ettan IPGphor User Manual 11003460 AE...

  • Page 14: Instructions

    3 Operation 3.1 Instructions Instructions Rehydrating the IPG strips Follow the instructions below to rehydrate the IPG strips. Step Action Rehydrate the IPG strips with the gel side down in the appropriate volume of rehydration solution using the Immobiline DryStrip Reswelling Tray. Rehydration in the Cup Loading Manifold is not recommended: the channel is too wide to ensure proper rehydration.
  • Page 15 Add the cover fluid evenly in the 12 manifold channels. Transfer the strips to the Ettan IPGphor Cup Loading Manifold. Place the strips under the cover fluid with the gel surface face up and the anodic (+, pointed) end to the anode of the Manifold.
  • Page 16 3 Operation 3.1 Instructions Step Action Center the strip down the length of the manifold channel. Protrusions along the sides guide the strip approximately straight, although some manual adjustment of the strip may be necessary, as shown below. 24 cm 18 cm 13 cm 11 cm...
  • Page 17 3 Operation 3.1 Instructions Step Action Place a strip of cups in the appropriate position as shown in the figure below, for example ~1 cm from the end of the gel portion of the IPG strip. Do NOT place the cup with the feet over a center protrusion. Push the cups into the channels with gloved fingers, starting at one end of the strip and working towards the other.
  • Page 18 3 Operation 3.1 Instructions Step Action Place the wicks on the IPG strips such that one end of the wick overlaps the end of the gel on the IPG strip, as shown in figure below. The electrode must contact the wick. With the electrode cams in the open position, place the electrode assembly on top of all the wicks.
  • Page 19 3 Operation 3.1 Instructions Sample application (loading) Follow the instructions below to load the sample. Step Action Briefly centrifuge the protein sample prior to loading to remove insoluble material and particulate matter. These materials will impede sample entry and result in vertical streaks in the second-dimension gel. Load samples into the sample cups.
  • Page 20 3 Operation 3.1 Instructions electrode assembly paper IPG strip wick hold down anode paper teeth bridge Ettan IPGphor User Manual 11003460 AE...

  • Page 21: Run Conditions

    3 Operation 3.2 Run conditions Run conditions Introduction Program the Ettan IPGphor with the recommended run parameters. Ramping the voltage slowly while the sample is entering the IPG strip will improve results. Instructions for optimal ramp, voltages and times, or Vhrs (volt-hours) totals are attached to each package of Immobiline Dry Strip.

  • Page 22: Maintenance

    4 Maintenance Maintenance Precautions WARNING Disconnect power. Always disconnect power from the instru- ment before performing any maintenance task. NOTICE The ceramic strip holders and manifold are very brittle and fragile. Take care in handling and do not subject to impacts. NOTICE Avoid scratching the electrode surfaces.
  • Page 23 4 Maintenance Manifold Light Clean the Ettan IPGphor Manifold Light with water, no warmer than 40 °C, and with IPG Strip Holder cleaning solution. NOTICE Do not use any organic solvents. Electrodes Wash the electrode assemblies with the Ettan IPGphor Strip Holder Cleaner. Rinse thoroughly with water then deionized water and allow to air dry.

  • Page 24: Troubleshooting

    5 Troubleshooting Troubleshooting Troubleshooting first-dimension IEF The following table specifies the troubleshooting situations that you might encounter while using the Ettan IPGphor Isoelectric Focusing System employing the Cup Loading Manifold. Symptom Possible cause Remedy Current is too low or zero Electrical continuity is Check the external elec- impeded.
  • Page 25 5 Troubleshooting Symptom Possible cause Remedy Check that the actual number of Immobiline DryStrip gels on the Ettan IPGphor platform is the same as the number of gels entered in the protocol. Conductivity/ionic Prepare the sample to strength is too high. yield a salt concentration less than 10 mM.
  • Page 26 5 Troubleshooting Symptom Possible cause Remedy Check that the feet of the cups are not resting on a centering protrusion in the channel. Incorrect strip place- Check that the strip is ment. centered inside of the channel. Sparking or burning in Current limit setting is Do not exceed the the Immobiline DryStrip...

  • Page 27: Reference Information

    6 Reference information Reference information Technical service and repair Cytiva offers complete technical support for all our products. If you have any questions about how to use this product, please contact your local Cytiva representative. Ordering information The following table provides you with information about replacement parts and their code numbers.
  • Page 28 © 2020–2021 Cytiva All goods and services are sold subject to the terms and conditions of sale of the supplying company operating within the Cytiva business. A copy of those terms and conditions is available on request. Contact your local Cytiva representative for the most current information.

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