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Pharmacia Biotech Multiphor II User Manual

Electrophoresis system
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Pharmacia Biotech Multiphor II User Manual

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Summary of Contents for Pharmacia Biotech Multiphor II

  • Page 1 (217) 352-9330 | Click HERE Find the GE Healthcare / Amersham Pharmacia Multiphor II Novablot at our website:...
  • Page 2 ¨ Multiphor Electrophoresis System User Manual 18-1103-43 Edition AF Artisan Technology Group - Quality Instrumentation ... Guaranteed | (888) 88-SOURCE | www.artisantg.com...
  • Page 3 Pharmacia Biotech AB. Meaning: Consult the insruction manual to avoid Pharmacia Biotech AB shall in no event be liable for persoanl injury, or damage to the product or other incidental or consequential damages, including without equipment.

  • Page 4: Table Of Contents

    5.10 Running conditions for precast gels......48 6. Maintenance................53 7. Technical specification.............55 8. Trouble shooting ..............57 9. Multiphor II application kits and accessories......61 9.1 SDS and Native PAGE, IEF Kit........61 9.2 Large Scale SDS and Native PAGE Kit ......65 9.3 Agarose IEF Kit ............66 9.4 Immunoelectrophoresis Kit..........69...
  • Page 5 9.15 Roller ................90 9.16 Sample application accessories ........91 9.17 Misellaneous .............93 10. Ordering information ............95 10.1 Multiphor II ..............95 10.2 MultiTemp III............99 10.3 EPS Power Supplies ...........99 10.4 Hoefer Automated Gel Stainer ........99 10.5 Precast gels and buffer strips ........100 10.6 Molecular weight and pl markers ......100...

  • Page 6: Introduction

    1. Introduction 1. Introduction Multiphor II electrophoresis system is a versatile modular system for horizontal electrophoresis, isoelectric focusing, 2-D electrophoresis and electrophoretic transfer. For ease of use and reproducible results, a unique range of precast gels for all major electrophoretic techniques is available with Multiphor II:...

  • Page 7: Electrophoretic Transfer

    1. Introduction The following guide summarizes how you can expand and use Multiphor II Electrophoresis Unit with application kits and accessories. Application Recommended Kit/Accessory Code No. SDS and Native PAGE SDS and Native PAGE, IEF Kit 18-1102-45 0.5x125x260 mm Gradient Maker...
  • Page 8 Multiphor II unit. 6. ‘‘Trouble shooting’’ offers suggestions for correcting problems that may occur. 7. ‘‘Multiphor II application kits and accessories’’ describes in detail the contents, assembly and use of each Multiphor II application kit. 8. ‘‘Ordering information.’’ - Multiphor II basic configurations, application kits,...
  • Page 9 1. Introduction Artisan Technology Group - Quality Instrumentation ... Guaranteed | (888) 88-SOURCE | www.artisantg.com...
  • Page 10 2. Safety information 2. Safety information To avoid any risk of injury, the instrument should be operated only by properly trained personnel and always in accordance with the instruction provided. Read this entire manual before using the instrument. WARNING! The instrument is designed for indoor use only. WARNING! Do not operate tthe system in extreme humidity (above 95% RH).
  • Page 11 2. Safety information WARNING! The cooling plate is rated for operation at up to 3.5 kV (p-p). Artisan Technology Group - Quality Instrumentation ... Guaranteed | (888) 88-SOURCE | www.artisantg.com...
  • Page 12 3. Description of parts 3. Description of parts This Multiphor II configuration includes; buffer tank with 4 levelling feet, 3.1 Multiphor II cooling plate with accessories, safety lid, electrode holder with movable Electrophoresis EPH/IEF electrodes (for buffer strips and electrode strips) and card-mounted Unit EPH electrodes (for electrophoresis using buffer chambers).
  • Page 13 3. Description of parts The larger pins are for connection to the safety lid and complete the electrical circuit when the lid is in position. The small left land pin is used to connect the EPH/IEF or card-mounted cathode electrodes. The small pin on the right connects to the card-mounted anode or the EPH/IEF cathode via the red lead mounted on the unit.
  • Page 14 NEVER EXCEED maximum pressure of 0.5 bar. The cooling tubing, tubing connector set and hose clamps provide a flexible and safe way to connect Multiphor II to a thermostatic circulator such as MultiTemp III. The electrode holder holds the movable EPH/IEF electrodes. The...
  • Page 15 3. Description of parts The electrode holder consists of a double strength glass plate with ground edges and four corner feet made of Rynite FR530. The electrode holder holds one anode and one cathode electrode. The EPH/IEF electrodes consist of moulded polysulfone bars which support the platinum wire, held taut by stainless steel springs.
  • Page 16 ProBind 45 NC 0.45 µm, roll 0.2x3.0 m 80-1247-86 GeneBind 45 nylon 0.45 µm, roll 0.2x3.0 m 80-1247-87 The Multiphor II buffer tank with the four adjustable levelling feet supports the NovaBlot electrodes. Artisan Technology Group - Quality Instrumentation ... Guaranteed | (888) 88-SOURCE | www.artisantg.com...
  • Page 17 WARNING! Polycarbonate is not resistant to concentrated acids and bases, or to halogenated and aromatic hydrocarbons. Precut electrode paper 200x250 mm and cellophane sheets, 210x320 mm, are included with Multiphor II NovaBlot Unit. Artisan Technology Group - Quality Instrumentation ... Guaranteed | (888) 88-SOURCE | www.artisantg.com...
  • Page 18 For easy connection of Multiphor II Electrophoresis Unit to the cooling device, install tubing connectors on both inlet and outlet tubings. Use one male and one female connector on the Multiphor II unit side and thermostat unit side respectively. The cooling plate tubings and thermostatic circulator tubings can then be locked separately.
  • Page 19 4. Installation To mount the EPH/IEF electrodes on the electrode holder, unscrew the clamping nut from each electrode. When running electrophoresis across the width of the cooling plate, mount the electrodes as illustrated. Place the electrode under the electrode holder. Replace the nut and lightly tighten until the electrode is held firmly in place.
  • Page 20 To align the electrodes with the electrode strips, place the electrode holder with the electrodes onto the false holes on the buffer tank using the four corner feet. Install the safety lid. 4.2 Multiphor II NovaBlot Unit for electro- phoretic transfer Multiphor buffer tank should be assembled with the four levelling feet.
  • Page 21 4. Installation The transfer sandwich can now be assembled on the bottom anode electrode. Full assembly instructions are given in Section 4.8. After assembling the transfer sandwich, saturate the upper cathode electrode with distilled water and remove any excess with absorbent paper. With the electrode lead to the front of the instrument, place the cathode electrode on top of the stack.

  • Page 22: Operation

    Electrophoresis in Practice, A Guide to Theory and Practice by Reiner Westermeier. Multiphor II contains two alternative electrode configurations. - EPH/IEF electrodes for use with buffer strips or electrode strips - EPH electrodes for use with electrode wicks and buffer chambers IEF, SDS-PAGE and native PAGE are most conveniently performed using EPH/IEF electrodes and buffer strips.

  • Page 23: Electrophoresis Using Excelgel Sds And Buffer Strips

    5. Operation 5.1 Electro- This section describes the running procedure for SDS PAGE using buffer strips. The running of ExcelGel SDS, gradient 8-18 using ExcelGel SDS phoresis buffer strips is chosen as an example, but the basic method is applicable to using all SDS PAGE and native PAGE gels.
  • Page 24 Samples are applied directly into the wells immediately prior to the run. Electrophoresis Connect Multiphor II to MultiTemp II thermostatic circulator. Switch on MultiTemp II 15 minutes before starting the experiment and set the temperature to 15 °C.
  • Page 25 5. Operation Place the electrode holder on the electrophoresis unit in the shallow depressions. Align the EPH/IEF electrodes with the centre of the buffer strips by loosening the clamping nuts and sliding the electrodes to the appropriate position. Retighten the clamping nuts. Lift the electrode holder slightly and reposition the supporting feet over the deep holes.
  • Page 26 Ending the run Turn off the power supply. Disconnect the Multiphor II unit from the power supply. Remove the safety lid from the unit. Carefully remove the electrode holder. Gently pull the strips from the gel and continue with detection techniques as required.
  • Page 27 5. Operation Detection For automated silver and coomassie staining of polyacrylamaida gels see Protocol guide, Hoefer Automated Gel Stainer (80-6343-34). Coomassie staining On completion of the electrophoresis, immediately immerse the gel in Staining Tray 1 using the solutions and times indicated in table below. The staining and destaining steps should be carried out on a shaking table.

  • Page 28: Electrophoresis Using Cleangel And Buffer Strips

    Ordering information and the information supplied with the CleanGel. Preparing the experiment Connect Multiphor II to MultiTemp II thermostatic circulator. Switch on MultiTemp II 20 minutes before starting the experiment and set the temperature to 15 °C. Note: Always wear clean gloves when working with polyacrylamide gels and buffer strips, particularly when using sensitive staining methods.
  • Page 29 5. Operation Rehydrate a CleanGel in an appropriate buffer using the GelPool, according to instructions supplied with the CleanGel. Pipette 1 ml kerosene onto the cooling plate. Position the gel in the middle of the cooling plate with the stiff plastic support film facing down. Position the sample wells at the cathodic or anodic side depending on the buffer system used.
  • Page 30 Apply 5-100 µl sample/well depending on gel type and sample concentration. Running conditions Place the electrode holder with the EPH/IEF electrodes on the Multiphor II unit. Align the electrodes so that they rest on the outer edges of the electrode strips.

  • Page 31: Electrophoresis Using Buffer Chambers

    5. Operation staining and preserving methods recommended for ExcelGel SDS (See Section 4.1) work well with CleanGel. This section describes the running procedure when using buffer chambers. 5.3 Electro- To place the electrodes in the buffer chamber, remove the cooling plate phoresis from the buffer tank.
  • Page 32 5. Operation do this, pour a few milliliters of kerosene or light paraffin oil towards one end of the cooling plate. Starting with one end of the gel support, gradually lower the gel to the horizontal position, constantly checking for trapped air bubbles. If air becomes trapped, raise the gel just enough to release the air and then continue to lower it onto the cooling plate.
  • Page 33 This will ensure even contact between the wicks and the gel and stop any moisture from condensing on the gel surface. Replace the safety lid on the unit and connect the Multiphor II unit to the power supply. Set the power requirements, and start the experiment.

  • Page 34: Isoelectric Focusing Using Ampholine Pagplate

    5. Operation 5.4 Isoelectric This section describes the running procedure for IEF using Ampholine PAGplate. For further information, see the instructions supplied with the focusing product. using Ampholine PAGplate Switch on MultiTemp II thermostatic circulator and set the temperature (normally 10 °C for polyacrylamide IEF) 15 minutes before starting the experiment.
  • Page 35 5. Operation Apply the electrode strips to the long edges of the gel. Make sure the electrode strips are applied with the correct polarity. Use sharp scissors to cut off the strips which protrude beyond the ends of the gel. Sample application Some proteins may be pH sensitive (they may precipitate at specific pH values) and better separation may be obtained when the sample is applied...
  • Page 36 5. Operation Starting the IEF run Place the electrode holder in the shallow depressions on the Multiphor II unit. Align the electrodes with the centre of the electrode strips by loosening the clamping nuts and sliding the electrodes to the appropriate position.
  • Page 37 Running conditions Connect Multiphor II to the power supply. Set the running conditions given in the table below. Start the isoelectric focusing by turning on the power supply. Observe the time limits closely. If the isoelectric focusing is run too long, the pH gradient will begin to drift towards the cathode.

  • Page 38: Isoelectric Focusing Using Cleangel Ief

    5. Operation Detection methods For automated silver and coomassie staining of polyacrylamaida gels see Protocol guide, Hoefer Automated Gel Stainer (80-6343-34). PhastGel Blue staining On completion of isoelectric focusing, remove the electrode strips from the gel. Immerse the gel in 250 ml of solution according to the schedule below (See section 4.9 for stock solutions).
  • Page 39 5. Operation Sample application Use the same methods as for Ampholine PAGplate (see Section 4.4). Running conditions In principle, CleanGel is run in the same way as Ampholine PAGplate except no electrode strips are used. The electrodes rest directly on the gel surface.

  • Page 40: Isoelectric Focusing Using Immobiline Dryplate

    5. Operation 5.6 Isoelectric Immobiline DryPlate is a dried polyacrylamide gel containing an immobilized pH gradient. focusing using Immobiline DryPlate Prior to use, the gel is rehydrated in distilled water or 1% Pharmalyte solution to a thickness of 0.5 mm in a special reswelling cassette. Additives such as urea and/or detergents can also be included at this stage.

  • Page 41: 2-D Electrophoresis Using Immobiline Drystrip

    Immobiline DryStrip for the first dimension of 2-D electrophoresis. The kit includes the accessories necessary to run up to 12 Immobiline DryStrip strips simultaneously on Multiphor II. The high sample capacity allows the application of up to 100 µl on each Immobiline DryStrip.
  • Page 42 5. Operation Immobiline DryStrip pH gradient shapes length (mm) of the strip pH gradient Shape Shape linear linear 3-10 linear linear 3-10 non-linear* *The pH gradient is flattened in the pH range 5-7 to achieve a high resolving area in this part of the gradient.

  • Page 43: Electrophoretic Transfer

    5. Operation Day 3: Second Dimension SDS-PAGE in ExcelGel gradient gels Activities: - Prepare equilibration solutions - Remove strips from 1st dimension - Equilibrate strips 2x10 minutes - Lay strips on 2nd dimension with SDS Markers - Run for approx. 3 hours - Add 1-2 hours for Coomassie staining and 4 hours for Silver staining.
  • Page 44 5. Operation Immobilizing membranes The immobilizing membrane is an important factor affecting the efficiency of the electrophoretic transfer. The most important requirements for an immobilizing membrane are: • High binding capacity for the molecules of interest • Preservation of the biologic activity of the molecules of interest •...
  • Page 45 5. Operation 1. Saturate the graphite anode plate with distilled water and remove the excess water with absorbent paper. With the electrode lead to the front of the instrument, fit the lower anode plate onto the buffer tank. Connect the anode socket (red lead) to the anode pin on the right side of the buffer tank.
  • Page 46 5. Operation 3. Carefully place the filter papers onto the anode electrode. When forming the first transfer sandwich, soak a further layer of three filter papers in anode solution S (see Section 4.9, Stock solutions), using the same method as above. Place them on top of the six filter papers on the anode electrode plate, again taking care to avoid trapping air bubbles.
  • Page 47 5. Operation 5. Loosen the support film from FilmRemover by pressing the handle and carefully lift the whole sandwich with the support film, immobilizing membrane and gel. Turn it over (support film up, immobilizing membrane down) and place it on the layer of three filter papers on the anode. Carefully remove the support film.
  • Page 48 Multiphor II base. 9. Close the Multiphor II safety lid and connect the unit to the power supply. It is recommended to run the transfer at a constant current of 0.8 mA/cm .
  • Page 49 5. Operation 10. When the transfer is complete, turn off the power supply and disconnect NovaBlot from the power supply. Remove the safety lid and the upper cathode electrode. Carefully disassemble the transfer sandwiches and remove the immobilizing membranes for analysis. If necessary, save and stain the gel to monitor the transfer efficiency.

  • Page 50: Stock Solutions

    5. Operation B. Sample buffer 5.9 Stock 0.050 mol/l Tris-HAc pH 7.5 solutions Dissolve 0.3 g Tris in 40 ml distilled water. Carefully adjust to pH 7.5 with HAc (approximately 0.14 ml). Make up to 50 ml with distilled water. Add 0.4 g SDS and a few grains of Bromophenol Blue.

  • Page 51: Running Conditions For Precast Gels

    5. Operation Transfer buffers using a discontinuous buffer system R. Anode solution 1, pH 10.4 Tris 36.3 g Methanol 200 ml Make up to 1000 ml with distilled water. S. Anode solution 2, pH 10.4 Tris 3.03 g Methanol 200 ml Make up to 1000 ml with distilled water.
  • Page 52 5. Operation CleanGel with Native Buffer Kit pH 8.9 Running conditions for CleanGel anodal electrophoresis with Native Buffer Kit, pH 8.9. Phase Voltage Current Power Time Temp (mA) (min) °C pre-run 300 * Or until the Bromophenol Blue front reaches the anodic wick. CleanGel with SDS Buffer Kit pH 8.0 Running conditions for CleanGel anodal electrophoresis with SDS Buffer Kit, pH 8.0.
  • Page 53 5. Operation CleanGel IEF 3-10 Running conditions for one CleanGel IEF 3-10. Phase Voltage Current Power Time Temp (mA) (min) °C Prefocusing Sample entry Isoelectric focusing 2000 Band sharpening 2500 If half a gel is used, halve the current and power settings. Immobiline DryPlate Running conditions for one Immobiline DryPlate.
  • Page 54 5. Operation Programme for Immobiline DryStrip, pH 3-10 NL, 180 mm. All steps are run at 10 °C. Phase Voltage (V) Current (mA) Power (W) Time (h) 2500 3500 10000 3500 32400 Total 19.5 44900* Programme for Immobiline DryStrip, pH 4-7, 110 mm. All steps are run at 10 °C.
  • Page 55 5. Operation Running conditions for Immobiline DryStrip, pH 4-7, 110 mm. Phase Voltage (V) Current (mA) Power (W) Time (h) 2200 13.5 29700 Running conditions for Immobiline DryStrip, pH 4-7, 180 mm. Phase Voltage (V) Current (mA) Power (W) Time (h) 3000 14.5 43500...
  • Page 56 5. Operation Artisan Technology Group - Quality Instrumentation ... Guaranteed | (888) 88-SOURCE | www.artisantg.com...
  • Page 57 5. Operation Artisan Technology Group - Quality Instrumentation ... Guaranteed | (888) 88-SOURCE | www.artisantg.com...
  • Page 58 5. Operation Artisan Technology Group - Quality Instrumentation ... Guaranteed | (888) 88-SOURCE | www.artisantg.com...

  • Page 59: Maintenance

    6. Maintenance 6. Maintenance A few standard measures are necessary to keep Multiphor II in full functioning order. After isoelectric focusing, remove the electrodes from the electrode holder and rinse with distilled water to remove the strong acidic and basic solutions.
  • Page 60 6. Maintenance Artisan Technology Group - Quality Instrumentation ... Guaranteed | (888) 88-SOURCE | www.artisantg.com...

  • Page 61: Technical Specification

    – used as a “stand alone” unit or connected to other CE labelled Pharmacia Biotech AB instruments or other products as recommended Artisan Technology Group - Quality Instrumentation ... Guaranteed | (888) 88-SOURCE | www.artisantg.com...
  • Page 62 7. Technical specifications Artisan Technology Group - Quality Instrumentation ... Guaranteed | (888) 88-SOURCE | www.artisantg.com...

  • Page 63: Trouble Shooting

    8. Trouble shooting 8. Trouble shooting WARNING! Turn OFF the power supply before opening the lid. Trouble shooting guide to PAGE Symptom Cause Remedy No current reading Safety plug improperly inserted Check the safety plug in power supply outlet insertion Pin and socket connection from Check the pin and socket electrode to base incomplete...
  • Page 64 8. Trouble shooting Symptom Cause Remedy Drying out of the gel Incorrect electrode solutions Use the recommended electrode near the electrodes solution at the specified concentration. Excessive power setting Check the power setting Sparking along edge Excess moisture on gel or under Remove the excess moisture of gel onto cooling plate cooling plate...
  • Page 65 8. Trouble shooting Symptom Cause Remedy Unevenly wetted electrode strips Electrode strips must be evenly wetted and be neither too wet nor too dry Electrode strips too short The strips should be cut just short of the edges of the gel Trouble shooting guide to electrophoretic transfer Symptom Cause...
  • Page 66 8. Trouble shooting Artisan Technology Group - Quality Instrumentation ... Guaranteed | (888) 88-SOURCE | www.artisantg.com...
  • Page 67 8. Trouble shooting Artisan Technology Group - Quality Instrumentation ... Guaranteed | (888) 88-SOURCE | www.artisantg.com...

  • Page 68: Multiphor Ii Application Kits And Accessories

    9. Multiphor II application kits and accessories 9. Multiphor II application kits and accessories This section describes the contents of the Multiphor II application kits and accessories and provides instructions for assembly and use. For experimental details including preparation of samples and stock solutions, running conditions, staining and preserving procedures see Chapter 4.
  • Page 69 9. Multiphor II application kits and accessories Optional accessories Designation Code No. Roller 80-1106-79 GelBond PAGfilm, 124x258 mm (50/pkg) 80-1129-36 Bind-Silane, 100 ml 17-1330-01 Repel-Silane, 500 ml 17-1331-01 Sample Application Syringe, 15 µl 80-1106-48 SDS Sample Application Strip, 26 samples, 40 µl...
  • Page 70 9. Multiphor II application kits and accessories To prevent the gel from sticking to the U-framed glass plate, coat the plate with Repel-Silane. Note: For this operation use gloves and a fume hood. Pour about 2 ml of Repel-Silane onto the glass plate and distribute it evenly with a tissue.
  • Page 71 9. Multiphor II application kits and accessories Form the mould by placing the U-framed glass plate in position and clamp together using four FlexiClamps. Note: Gloves must be worn to protect the user from contact with the toxic acrylamide solution.
  • Page 72 9. Multiphor II application kits and accessories 9.2 Large Scale This kit is used for casting 0.5 mm homogeneous or gradient polyacrylamide gels. The gels are cast only on GelBond PAGfilm. The kit is recommended SDS and for casting large format gels for 2-D electrophoresis.

  • Page 73: Agarose Ief Kit

    9. Multiphor II application kits and accessories Gel Knife 80-1106-37 Blades (5/pkg) 80-1106-38 Template, 125x260 mm (10/pkg) 80-1129-55 Staining Tray 2, 60x260x320 mm 18-1018-09 Cellophane Sheets (50/pkg) 80-1129-38 The 4 mm glass plate is used as a support for GelBond PAGfilm.
  • Page 74 9. Multiphor II application kits and accessories Optional accessories Designation Code No. Roller 80-1106-79 GelBond film, 124x258 mm (50/pkg) 80-1129-32 Sample Application Syringe, 15 µl 80-1106-48 EPH/IEF Sample Application Foil, 24 samples 2-4 µl (5/pkg) 80-1129-47 Humidity Chamber 125x260 mm (4/pkg) 18-1016-78 The 3 mm thick glass plate is used as a support for GelBond film.
  • Page 75 9. Multiphor II application kits and accessories Prepare the agarose solution in a water bath and cool to 75 °C before adding the carrier ampholytes. Take the mould out of the heating cabinet. Slowly fill a 20 ml syringe (with flexible tubing attached) with the molten agarose solution.

  • Page 76: Immunoelectrophoresis Kit

    9. Multiphor II application kits and accessories 9.4 Immuno- This kit is used for casting agarose gels on a 84x94 mm glass plate or GelBond film for immunoelectrophoresis or immunodiffusion. Templates are electro- included for the following techniques phoresis Kit •...
  • Page 77 9. Multiphor II application kits and accessories Gel Knife 80-1106-37 Blade (5/pkg) 80-1106-38 Tape, Dymo 0,25x9 mm, 3 m 80-1129-50 Electrophoresis Wick, 82x130 mm (500/pkg) 80-1129-53 Tygon Tubing, 2.4/4.0 mm, 2 m 80-1006-70 Adaptor Connector 80-1011-88 Optional accessories Designation Code No.
  • Page 78 9. Multiphor II application kits and accessories Leave the gels to solidify for l5 minutes at room temperature. For the best results, store the gels in Humidity Chamber at 4 °C for one hour to allow complete solidification. Gels may be stored in this manner for several days.
  • Page 79 9. Multiphor II application kits and accessories Using the Sample Application Syringe, carefully fill the wells making sure not to damage the gel. For immunoelectrophoretic techniques, begin the electrophoresis as soon as possible to minimize diffusion. Diffusion techniques can be performed in the Humidity Chamber.

  • Page 80: Preparative Ief Kit

    9. Multiphor II application kits and accessories 9.5 Preparative This kit is used for preparative IEF in a granulated gel, Ultrodex. The separation is performed in a tray measuring 125x260x5 mm. IEF Kit The maximum capacity of the tray is 150 ml of initial Ultrodex gel slurry.
  • Page 81 Place the prepared tray containing gel onto the cooling plate (cooled to 10 ° C) of Multiphor II. A film of water containing a non-ionic detergent (e.g. 0.1% Triton X-100) is used to aid thermal conductivity.
  • Page 82 9. Multiphor II application kits and accessories Place IEF Electrode Strips soaked in the appropriate electrode solution at the anodic and cathodic side, each on top of the strips already placed in the tray. Cut off the protruding parts so that the strips fit exactly into the tray. If desired, prefocus the gel.
  • Page 83 9. Multiphor II application kits and accessories Stain the entire paper print or, if required, cut it into parallel strips for treatment with different staining techniques (e.g. general protein stain, zymogram stain or glycoprotein stain). If the staining procedure is time-consuming, reapply the power to the gel to avoid diffusion or insert the fractionating grid frame with blades immediately after the paper print has been taken.

  • Page 84: Immobiline Drystrip Kit

    9. Multiphor II application kits and accessories Using a spatula, transfer each gel section to a PEGG Elution Column. Add just enough of a suitable elution buffer to each column to resuspend the gel, and let the mixture settle until all of the buffer has entered the gel bed. Then, carefully add one gel volume of buffer and continue the elution.

  • Page 85: Novablot Kit

    9. Multiphor II application kits and accessories 9.7 NovaBlot Kit This kit is used for electrophoretic transfer of proteins from polyacrylamide or agarose gels to an immobilizing membrane. The maximum gel size is 200x250 mm. By building transfer sandwiches, simultaneous transfer from several gels of the same type can be achieved.

  • Page 86: Ultromould

    9. Multiphor II application kits and accessories 9.8 UltroMould UltroMould provides a simple and reliable method for casting ultrathin polyacrylamide gels with a size of 125x258 mm. Using the patented sliding plate technique, gels with a uniform thickness of 0.1, 0.2, 0.3, 0.4 or 0.5 mm can be prepared.
  • Page 87 9. Multiphor II application kits and accessories Place the Gel Casting Table in a fume cupboard. (It is very important to work in a fume cupboard as acrylamide and bis are neurotoxins). Place the clean long glass plate on the lower shelf of the Gel Casting Table so that one end touches the white end screw.
  • Page 88 9. Multiphor II application kits and accessories Making sure that the gel spacer is completely flat, attach the four holding guides to the rails along the sides of the Gel Casting Table. The flat side holds the spacer securely in place and guides the movement of the short glass plate when filling the cassette.
  • Page 89 9. Multiphor II application kits and accessories Before disassembling the mould, carefully remove any unpolymerized gel solution. Remove the weight, the holding guides and the black end screw. To remove the gel, insert a spatula under the gel spacer and between the film and the long glass plate and twist slightly to separate them.

  • Page 90: Gradient Maker

    9. Multiphor II application kits and accessories 9.9 Gradient Maker This Gradient Maker comprises two compartments, the mixing chamber, closest to the outlet, and the reservoir. The Compensating Stick is used for linear gradients and corrects for the density difference between the light and dense solutions and the volume of the magnetic stirrer.
  • Page 91 9. Multiphor II application kits and accessories If the channel is blocked with polymerized gel solution, disassemble and clean the Gradient Maker as described below. Close the channel by pushing the valve to closed (c) position. Check that the pinch cock is closed.

  • Page 92: Reswelling Cassette

    9. Multiphor II application kits and accessories Open the valve between the chambers and then the pinch cock on the outlet tubing to begin gradient formation. Check that the solution begins to flow into the mould. When the mould has been filled, rinse the Gradient Maker with water to avoid polymerization of remaining gel solution in the channel.
  • Page 93 9. Multiphor II application kits and accessories Cut a 10 cm length of silicone tubing and make a 2 cm long cut across one end of the tubing. Note: It is important that the cut is 2 cm long since it may be difficult to get the tubing through the hole in the glass plate with a shorter cut.
  • Page 94 9. Multiphor II application kits and accessories gel surface as this may start rehydration and prevent proper rehydration of the entire gel. Sticky spots on the gel will form lead to bubble formation during addition of rehydrating solution. To prevent the gel from adhering to the U-framed glass plate, coat the plate with Repel-Silane.

  • Page 95: Levelling Set

    9. Multiphor II application kits and accessories When opening the Reswelling Cassette, first remove the 3 mm thick glass plate and then separate the gel and the U-framed glass plate. Check the rehydrated gel and remove excess rehydration solution with a paper tissue.

  • Page 96: Filmremover

    9. Multiphor II application kits and accessories FilmRemover is used for removing backing from a gel before electrophoretic 9.13 FilmRemover transfer. Polyacrylamide or agarose gels with a thickness between 0.1 mm and 5.0 mm and a maximum gel size of 200x245 mm can be used.

  • Page 97: Staining Tray 1 And 2

    9. Multiphor II application kits and accessories 9.14 Staining Tray Both trays have lids with a recessed handle allowing several trays to be stacked. 1 and 2 Please note that only Staining Tray 1 has a removable gel holder. Designation Code No.

  • Page 98: Sample Application Accessories

    9. Multiphor II application kits and accessories Several accessories are available for simple and convenient sample 9.16 Sample application with Multiphor II. application accessories Designation Code No. Sample Application Syringe, 15 µl 80-1106-48 The glass syringe accurately dispenses up to 15 µl of fluid and is supplied with a short needle for convenient sample application.
  • Page 99 9. Multiphor II application kits and accessories Up to 52 samples can be applied with this strip. Each well holds up to 20 µl of sample. The applicator strip is made of flexible silicone and is applied directly onto the gel surface.
  • Page 100 9. Multiphor II application kits and accessories Designation Code No. EPH/IEF Sample Application Foil 24 samples, 2-4 µl 80-1129-47 This application foil with narrow slits is recommended for electrophoresis and IEF in agarose gels. Up to 24 samples can be applied, with a sample volume of 2-4 µl in each slit.
  • Page 101 9. Multiphor II application kits and accessories Artisan Technology Group - Quality Instrumentation ... Guaranteed | (888) 88-SOURCE | www.artisantg.com...

  • Page 102: Ordering Information

    10. Ordering information 10. Ordering information WARNING! Only spare parts approved or supplied by Pharmacia Biotech AB may be used for maintaining and servicing of MULTIPHOR II Product Quantity Code No. 10.1 Multiphor II Basic configuration Multiphor II 18-1018-06 Electrophoresis Unit...
  • Page 103 10. Ordering information Product Quantity Code No. Staining Tray 2 18-1018-09 with lid, no gel holder, 60x260x320 mm Roller 80-1106-79 Template, 125x260 mm 80-1129-55 PEGG Elution Columns 80-1106-75 Gel Knife (no blades) 80-1106-37 Blades (for Gel Knife) 80-1106-38 Gel Puncher, 2.0 mm 80-1109-69 Gel Puncher, 2.5 mm 80-1109-70...
  • Page 104 10. Ordering information Product Quantity Code No. Electrodes and electrode holders EPH/IEF Electrode, anode 18-1121-53 EPH/IEF Electrode, cathode 18-1121-52 Electrode Holder (for 18-1106-60/-61) 80-1106-55 EPH Electrode (long), anode 18-1122-20 EPH Electrode (long), cathode 18-1122-19 Electrode (Immobiline DryStrip Kit), anode 18-1018-66 Electrode (Immobiline DryStrip Kit), cathode 18-1018-67...
  • Page 105 10. Ordering information Product Quantity Code No. 125x270x6 mm (UltroMould) 80-1115-81 125x490x6 mm (UltroMould) 80-1115-80 Glass plate treatment Bind-Silane 100 ml 17-1330-01 Repel-Silane 500 ml 17-1331-01 Gel support GelBond PAGfilm, 124x258 mm 80-1129-36 203x260 mm 80-1129-37 GelBond Film (agarose), 84x94 mm 80-1129-33 124x258 mm 80-1129-32...

  • Page 106: Hoefer Automated Gel Stainer

    10. Ordering information Product Quantity Code No. Membranes, electrophoretic transfer Nitrocellulose, 0.20 µm, 150x200 mm 80-1098-91 Nitrocellulose, 0.45 µm, 150x200 mm 80-1098-90 ProBind 45 NC 0.45 µm, roll 0.2x3.0 m 80-1247-86 GeneBind 45 nylon 0.45 µm, roll 0.2x3.0 m 80-1247-87 Tapes Dymo, 0.25x9 mm, 3 m 80-1129-50...

  • Page 107: Precast Gels And Buffer Strips

    10. Ordering information Product Quantity Code No. SDS-PAGE and Native PAGE 10.5 Precast gels ExcelGel SDS Homogeneous 7.5 80-1260-01 and buffer ExcelGel SDS Homogeneous 12.5 80-1261-01 ExcelGel SDS Homogeneous 15 80-1262-01 strips ExcelGel SDS, gradient 8-18 80-1255-53 ExcelGel XL SDS, gradient 12-14 17-1236-01 ExcelGel SDS Buffer Strips anode and cathode...

  • Page 108: Molecular Weight And Pl Markers

    10. Ordering information Product Quantity Code No. Second dimension ExcelGel SDS, gradient 8-18 80-1255-53 110x245x0.5 mm ExcelGel XL SDS, gradient 12-14 17-1236-01 180x245x0.5 mm ExcelGel SDS Buffer Strips anode and cathode 6 each 17-1342-01 MW range 2.512-16.949, 2 mg 80-1129-83 10.6 Molecular MW range 14.000-94.000, 200 µg/vial 17-0446-01...

  • Page 109: Agarose, Ultrodex And Plusone Electrophoresis Chemicals

    10. Ordering information Product Size Code No. Immobiline Immobiline II pK 3.6 10 ml 80-1255-70 Immobiline II pK 4.6 10 ml 80-1255-71 Immobiline II pK 6.2 10 ml 80-1255-72 Immobiline II pK 7.0 10 ml 80-1255-73 Immobiline II pK 8.5 10 ml 80-1255-74 Immobiline II pK 9.3...
  • Page 110 10. Ordering information PlusOne electrophoresis chemicals Product Quantity Storage Code No. Gel casting chemicals Acrylamide IEF IEF, PAGE, Sequencing 250 g 17-1300-01 Acrylamide IEF IEF, PAGE, Sequencing 1000 g 17-1300-02 Acrylamide IEF 40% solution IEF, PAGE 1000 ml 17-1301-01 ReadyMix IEF 41.5 g1 17-1309-01 ReadySol IEF T40 C3...
  • Page 111 10. Ordering information Artisan Technology Group - Quality Instrumentation ... Guaranteed | (888) 88-SOURCE | www.artisantg.com...
  • Page 112 10. Ordering information Artisan Technology Group - Quality Instrumentation ... Guaranteed | (888) 88-SOURCE | www.artisantg.com...
  • Page 113 Artisan Technology Group - Quality Instrumentation ... Guaranteed | (888) 88-SOURCE | www.artisantg.com...

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