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GE Ettan IPGphor 3, Operating Instructions Manual

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Action

Step

Pipette the solution slowly at a central point in the strip holder channel away
from the sample application wells.

1

Remove any larger air bubbles.

2

50

Ettan IPGphor 3 Operating Instructions 28-9635-51 AC

5 Operation

5.1 Preparations before start
5.1.1 Use the strip holders

Summary of Contents for Ettan IPGphor 3

Page 1: ...Ettan IPGphor 3 Operating Instructions Original instructions ...

Page 2: ...Page intentionally left blank ...

Page 3: ...g and transport 41 4 3 Setup 45 4 4 Software installation for isoelectric focusing process 46 5 Operation 47 5 1 Preparations before start 48 5 1 1 Use the strip holders 59 5 1 2 Use the Ettan IPGphor Cup Loading Manifold 66 5 2 Start up the instrument 68 5 3 Perform a run from the control panel 69 5 3 1 Ettan IPGphor 3 control panel functionality 73 5 3 2 Protocol parameters 75 5 3 3 Set protocol...

Page 4: ...ing 111 8 Reference information 112 8 1 Health and Safety Declaration Form 114 8 2 Technical specifications 116 8 3 Ordering information 118 Index 4 Ettan IPGphor 3 Operating Instructions 28 9635 51 AC Table of Contents ...

Page 5: ...ortant user infor mation regulatory information and lists of associated documentation In this chapter See page Section 6 1 1 About this manual 7 1 2 Important user information 9 1 3 Regulatory information 12 1 4 Associated documentation Ettan IPGphor 3 Operating Instructions 28 9635 51 AC 5 1 Introduction ...

Page 6: ...des you with the instructions needed to use the Ettan IPGphor 3 Isoelectric Focusing System together with Ettan IPGphor 3 Typographical conventions Software items are identified in the text by bold italic text A colon separates menu levels thus File Open refers to the Open command in the File menu Hardware items are identified in the text by bold text e g Power switch 6 Ettan IPGphor 3 Operating I...

Page 7: ...lectrophoresis The system comprises GE s Ettan IPGphor 3 combined with separate strip holders or a manifold Two types of manifolds are available the ceramic manifold and the light manifold The IEF separation protocols are programmed run and controlled from the Ettan IPGphor 3 control panel or from a PC running the Ettan IPGphor 3 Control Software Ettan IPGphor 3 is intended to be used as a laborat...

Page 8: ...ous situation which if not avoided could result in minor or moderate injury It is important not to pro ceed until all stated conditions are met and clearly understood Notices NOTICE NOTICE indicates instructions that must be followed to avoid damage to the product or other equipment Notes and tips A Note is used to indicate information that is important for trouble free and optimal use of the prod...

Page 9: ...ent Content Requirement GE Healthcare Bio Sciences AB Name and address of manufacturer Björkgatan 30 SE 751 84 Uppsala Sweden Conformity with EU Directives This product complies with the European directives listed in the table by fulfilling the corresponding harmonized standards Title Directive Machinery Directive MD 2006 42 EC Electromagnetic Compatibility EMC Directive 2004 108 EC Low Voltage Di...

Page 10: ...C Electrical equipment for measurement controland laboratory use EMC re quirements EN IEC 61326 1 Emission according to CISPR 11 Group 1 class A EN ISO standard is harmo nized with EU directive 2006 42 EC Safety of machinery Gen eral principles for design Risk assessment and risk reduction EN ISO 12100 FCC compliance This device complies with part 15 of the FCC Rules Operation is subject to the fo...

Page 11: ...e Regulatory compliance of connected equipment Any equipment connected to Ettan IPGphor 3 Isoelectric Focusing System should meet the safety requirements of EN 61010 1 IEC 61010 1 or relevant harmonized standards Within the European Union connected equipment must be CE marked Environmental conformity This product conforms to the following environmental requirements Title Requirement Restriction of...

Page 12: ...or Cup Loading Manifold User Manual contains instructions for first di mension isoelectric focusing of proteins on IPG strips Ettan IPGphor 3 Control Software User Manual 2 D Electrophoresis principles and methods guide Instructions attached to each package of Immobiline DryStrip 12 Ettan IPGphor 3 Operating Instructions 28 9635 51 AC 1 Introduction 1 4 Associated documentation ...

Page 13: ...c Focusing System The labels on the system and information regarding recycling are also described In this chapter See page Section 14 2 1 Safety precautions 20 2 2 Labels 22 2 3 Emergency procedures 24 2 4 Recycling information 25 2 5 Declaration of Hazardous Substances DoHS Ettan IPGphor 3 Operating Instructions 28 9635 51 AC 13 2 Safety instructions ...

Page 14: ...sks due to the process or pro cess environment Evaluate the effect the system and the processes where it is used have to the classification of the hazardous area The process might cause the area to increase or the zone classifi cation to change Implement the risk reduction measures needed including use of personal protection equipment WARNING The customer must make sure that all installation maint...

Page 15: ...ways use appropriate personal protective equipment during in stallation operation inspection maintenance and cleaning of Ettan IPGphor 3 WARNING When using hazardous chemical and biological agents take all suitable protective measures such as wearing protective glasses masks and gloves resistant to the substances used and protect self from hazardous substances that are used Follow local and or nat...

Page 16: ...ke appropriate measures Using flammable liquids WARNING Before starting the system make sure that there is no leakage WARNING A fume hood or similar ventilation system shall be installed when flammable or noxious substances are used Installation WARNING Make sure that the supply voltage at the wall outlet corresponds to the marking on the instrument before connecting the power cord WARNING The Ett...

Page 17: ...em operation WARNING The Ettan IPGphor 3 is capable of producing thousands of volts Before operating the unit read and fully understand the Ettan IPGphor 3 operating instructions and warnings WARNING Check that the high voltage is turned off before opening the safety lid The HV ON indicator on the front panel should not light WARNING Ettan IPGphor 3 must never be used if any safety functions are o...

Page 18: ...aratus to function correctly There should be a minimum of 10 cm unobstructed space behind the rear panel of the instrument WARNING Avoid spilling liquids on the body of the instrument If large volumes of liquid have penetrated the casing of the instrument and come into contact with the electrical components immediately switch off the instrument and contact an authorized service technician CAUTION ...

Page 19: ...n give a lethal electric shock Service and planned maintenance should be performed by personnel au thorized by GE WARNING Do not remove the main cover There are no userserviceable components inside and you can be exposed to high voltage WARNING Always disconnect power from the instrument and detach the power cord before replacing fuses CAUTION Do not use any organic solvents during cleaning Ettan ...

Page 20: ...XX VA Fuse XX X XXAL 250 V Made in Sweden GE Healthcare Bio Sciences AB 751 84 Uppsala Sweden Protection Class XXXX Conforms to ANSI UL Std 61010 1 Cert to CAN CSA Std C22 2 No 61010 1 4001767 System label description Description Label Name of product Product name Instrument Assembly Number Code no Instrument serial number Serial no Manufacturing year Mfg Year Voltage Voltage Frequency Frequency M...

Page 21: ...es with applicable European directives Refer to Conformity with EU Directives on page 9 The system complies with the requirements for electromagnetic compliance EMC in Australia and New Zealand This symbol indicates that Ettan IPGphor has been certified by a Nationally Recognized Testing Laboratory NRTL NRTL means an organization which is recognized by the US Occupational Safety and Health Adminis...

Page 22: ...en if an uninterruptible power supply UPS is used 1 2 Power failure In the event of system shutdown due to power failure emergency stop or process inter ruption malfunctions must be rectified before the Ettan IPGphor 3 is restarted The result of a power failure depends on which unit is affected will result in Power failure to Any experiments that are running will stop The time for the stop will be...

Page 23: ...cy shutdown power failure When the power returns to normal no experiments that were running at the time of emergency shutdown will be continued Proceed as follows Action Step Start the instrument see Turn on power and run diagnostic program on page 66 1 Restart the software 2 When the system is connected the log event will be stored in the system log If you wish to continue a run set up and run a ...

Page 24: ...and recycled according to national and local environmental regulations Recycling of hazardous substances Ettan IPGphor 3 contains hazardous substances Detailed information is available from your GE representative Disposal of electrical components Waste of electrical and electronic equipment must not be disposed as unsorted municipal waste and must be collected separately Please contact an authoriz...

Page 25: ...riod EFUP which indicates the period during which the toxic or hazardous substances or elements contained in electronic information products will not leak or mutate under normal operating con ditions so that the use of such electronic information products will not result in any severe environmental pollution any bodily injury or damage to any assets The unit of the period is Year In order to maint...

Page 26: ...列数据为发布时所能获得的最佳信息 0 Indicates that this toxic or hazardous substance contained in all of the homogeneous ma terials for this part is below the limit requirement in SJ T11363 2006 X Indicates that this toxic or hazardous substance contained in at least one of the homoge neous materials used for this part is above the limit requirement in SJ T11363 2006 Data listed in the table represents best inform...

Page 27: ... and an overview of its components In this chapter This chapter contains the following sections See page Section 28 3 1 Illustrations of Ettan IPGphor 3 32 3 2 Ettan IPGphor 3 instrument 34 3 3 Ettan IPGphor 3 Control Software 35 3 4 Strip holders Ettan IPGphor 3 Operating Instructions 28 9635 51 AC 27 3 System description ...

Page 28: ...r a manifold Two types of manifolds are available the ceramic manifold and the light manifold Ettan IPGphor 3 Isoelectric Focusing System also includes Immobiline DryStrip gel strips which contain an immobilized pH gradient IPG For more information about the manifolds see Ettan IPGphor Cup Loading Manifold User Manual Note 28 Ettan IPGphor 3 Operating Instructions 28 9635 51 AC 3 System descriptio...

Page 29: ...e electrod area Control panel Safety lid 1 2 3 4 Function Part Safety lid 1 Positive anode electrode area 2 Negative cathode electrode area 3 Control panel 4 Ettan IPGphor 3 Operating Instructions 28 9635 51 AC 29 3 System description 3 1 Illustrations of Ettan IPGphor 3 ...

Page 30: ...he LCD screen 1 3 4 5 6 7 8 9 2 Function Part START button 1 STOP button 2 LCD screen Main screen 3 Left arrow button 4 Up arrow button 5 Right arrow button 6 Down arrow button 7 EDIT button 8 HV ON indicator lamp 9 30 Ettan IPGphor 3 Operating Instructions 28 9635 51 AC 3 System description 3 1 Illustrations of Ettan IPGphor 3 ...

Page 31: ...s are placed on the rear panel of the Ettan IPGphor 3 1 2 3 4 Function Part Power switch 1 Fuse holder 2 RS232 for connection to computer 3 Power inlet 4 Ettan IPGphor 3 Operating Instructions 28 9635 51 AC 31 3 System description 3 1 Illustrations of Ettan IPGphor 3 ...

Page 32: ...tric modules Both electrode areas are gold plated copper Placing a manifold or a strip holder so that it bridges both electrode areas completes the electrical circuit once the safety lid is in position Protective cover When analyzing proteins labeled with light sensitive stains like CyDye a light protective cover is used The cover is applied by magnetic force but is easy to partially remove for ch...

Page 33: ...hor 3 standard strip holders a lid adapter must be used to apply the correct amount of pressure to the lids of the Ettan IPGphor 3 strip holders Ettan IPGphor 3 Operating Instructions 28 9635 51 AC 33 3 System description 3 2 Ettan IPGphor 3 instrument ...

Page 34: ...3 Control Software controls up to four Ettan IPGphor 3 systems at one time each running a different set of run parameters The software allows programming and recommended protocols are generated by pro viding instrument configuration IPG strip length and pH gradient The software records the run parameters over time and presents data as graphs and log files Data is saved or can be exported to Micros...

Page 35: ... fragile and must be handled with care The ceramic is treated with a special coating to minimize protein binding and must be cleaned only with the Ettan IPGphor Cleaning Solution supplied to retain that coating Each fixed length strip holder base holds a single IPG strip throughout rehydration and isoelectric focusing A matching length holder is available for each size strip All strips and holders...

Page 36: ... electrode while allowing electrolysis gases to escape Rehydration and sep aration progress can be monitored visually through the clear cover Cover Strip holder cover The cover is made of acrylic and protects the IPG strip during rehydration The shape of the underside of the cover ensures electrical continuity during IEF by pressing the strip against the electrode while allowing electrolysis gases...

Page 37: ...Isoelectric Focusing System In this chapter This chapter contains the following sections See page Section 38 4 1 Site requirements 40 4 2 Unpacking and transport 41 4 3 Setup 45 4 4 Software installation for isoelectric focusing process Ettan IPGphor 3 Operating Instructions 28 9635 51 AC 37 4 Installation ...

Page 38: ... A Noise emission 1 High relative humidity may result in condensation forming on the platform surface Runs con ducted at a platform temperature of 20 C and relative humidity less than 70 will generally not cause condensation Platform temperatures greater than 20 C allow for higher relative humidity without condensation Environments for which condensation occurs on the platform surface may vary If ...

Page 39: ...n Parameter 100 240 V AC 10 Mains supply voltage 50 60 Hz Frequency 230 VA Power consumption Overvoltage category II Transient overvoltages Ettan IPGphor 3 Operating Instructions 28 9635 51 AC 39 4 Installation 4 1 Site requirements ...

Page 40: ...diately Please keep all packing material and use if damaged parts need to be returned for repair or replacement Check the equipment for damage before starting assembly and installation Document any damage and contact your local GE Remove straps and packing material and stand equipment upright before starting installation Transport Before moving the system Disconnect all cables Lift the instrument ...

Page 41: ... power switch on the rear panel of the instrument 1 Position the instrument on a flat surface with the spirit level in the center Turn the leveling feet as necessary to make the instrument level and stable 2 Power connection WARNING Make sure that the supply voltage at the wall outlet corresponds to the marking on the instrument before connecting the power cord WARNING Only use power cords deliver...

Page 42: ...nded power outlet 2 Turn on power and run diagnostic program Turn on the instrument and run a diagnostic program according to the description in Section 5 2 Start up the instrument on page 66 Serial port connector for a computer connection NOTICE Any computerused with the equipmentshall comply with IEC 60950 and be installed and used according to the manufacturer s instruc tions The RS232 serial p...

Page 43: ... Data bits 1 Stop bit 1 Start bit None Parity None Flow control Set the baud rate Follow the instructions below to set the baud rate on the LCD screen Action Step Press the Up and Down arrows together to access the Baud menu 1 Use the Up or Down arrows to set the baud rate 2 Use the Right arrow to exit to the main menu 3 S e r i a l P o r t S e t u p B a u d R a t e 9 6 0 0 S e t U p o r D n E x i...

Page 44: ...re it can be used Strip holder or manifold Strips Buffer and sample liquids Spare parts and accessories For correct up to date information on spare parts and accessories visit www gelifesciences com 44 Ettan IPGphor 3 Operating Instructions 28 9635 51 AC 4 Installation 4 3 Setup ...

Page 45: ...process Introduction Refer to Ettan IPGphor 3 Control Software User Manual to install the Ettan IPGphor 3 Control Software Ettan IPGphor 3 Operating Instructions 28 9635 51 AC 45 4 Installation 4 4 Software installation for isoelectric focusing process ...

Page 46: ... the method selected either fixed length Ettan IPGphor 3 strip holders or use the Ettan IPGphor Cup Loading Manifold In this chapter This chapter contains the following sections See page Section 47 5 1 Preparations before start 66 5 2 Start up the instrument 68 5 3 Perform a run from the control panel 90 5 4 Run a pre programmed protocol from the PC 46 Ettan IPGphor 3 Operating Instructions 28 963...

Page 47: ...urea may be warmed briefly Do not heat any solutions containing urea above 30ºC to 40ºC as isocyanate a urea degradation product will carbamylate the proteins in the sample thus changing their isoelectric points NOTICE All chemicals should be of the highest purity Double distilled water should be used Ettan IPGphor 3 Operating Instructions 28 9635 51 AC 47 5 Operation 5 1 Preparations before start...

Page 48: ...ld not light Prepare the strip holder s NOTICE Handle the ceramic holders with care as they are brittle and fragile Action Step Select the strip holder s corresponding to the IPG strip length chosen for the experiment 7 11 13 18 or 24 cm 1 Wash each holder with the strip holder cleaning solution supplied to remove residual protein 2 Rinse thoroughly with double distilled water 3 Use a cotton swab ...

Page 49: ...ted the casing of the instrument and come into contact with the electrical components immediately switch off the instrument and contact an authorized service technician CAUTION Always wear gloves when handling IPG strips and the equipment that comes in contact with them This will help minimize protein contamination which can result in artifactual spots in stained 2 D gel patterns NOTICE Use the ap...

Page 50: ... a central point in the strip holder channel away from the sample application wells 1 Remove any larger air bubbles 2 50 Ettan IPGphor 3 Operating Instructions 28 9635 51 AC 5 Operation 5 1 Preparations before start 5 1 1 Use the strip holders ...

Page 51: ...and lower the strip and slide it back and forth along the surface of the solution tilting the strip holder slightly as needed to assure complete and even wetting 3 Lower the cathodic square end of the strip into the channel making sure that the IPG gel is in contact with the strip holder electrodes at each end The gel can be visually identified once the rehydration solution begins to dye the gel B...

Page 52: ...n a level surface A minimum of 10 h is required for rehydration overnight is recommended Alternatively the rehydration period can be programmed as the first step of an Ettan IPGphor 3 protocol This is especially convenient if temperature control during re hydration is a concern or if a low voltage is applied during rehydration Sample rehydration under voltage optional Follow the instructions below...

Page 53: ... Follow the instructions below to apply electrode pads prior to IEF Action Step Cut two 3 mm wide electrode pads from a paper IEF electrode strip 18 1004 40 1 Position on a clean flat surface such as a glass plate and soak with deionized water Remove excess water by blotting with tissue paper Note Electrode pads must be damp not saturated or dripping 2 Lift one end of the rehydrated IPG strip usin...

Page 54: ...rip Note Action Step Prepare the sample in a solution similar in composition to the rehydration solution used 1 Pipet the sample into either or both of the lateral wells at either end of the strip holder Introduce the sample below the cover fluid Up to 7 5 μl of sample solution can be added to each side i e 15 μl per well or maximum 30 μl if both sides of both wells are used 2 54 Ettan IPGphor 3 O...

Page 55: ... holder 2 is over the anode pointing to the back of the unit and the blunt end is over the cathode Guidemarks along the sides of the platform show approximate positioning for each strip holder size 7 11 13 18 and 24 cm The pointed end of the strip holder is in the anodic electrodae area Electrode contacts underneath one in each electrode area Anodic electrode area Cathodic electrode area 1 2 4 3 E...

Page 56: ...older electrodes and the electrode areas damaging the instrument When using the Ettan IPGphor standard strip holders lid adapters must be used to apply the correct amount of pressure to the lids of the IPGphor strip holders The pressure keeps the IPG strip in contact with the IPG strip holder electrodes 56 Ettan IPGphor 3 Operating Instructions 28 9635 51 AC 5 Operation 5 1 Preparations before sta...

Page 57: ...p Apply two lid adapters 2 right across the lids of the strip holders one over the anodic area and the other over the cathodic area 1 1 2 3 Use two or more strip holders 3 and place them wide apart to ensure that the lid adapters are only hanging on one strip holder and not pressing down on the other 2 Carefully close the safety lid 1 by applying light downwards pressure ensure the mechanism locks...

Page 58: ...ping the run 4 Light prot cover To open the safety lid press down the lid and the lock is released and the run is stopped Close the lid and the run continues 5 58 Ettan IPGphor 3 Operating Instructions 28 9635 51 AC 5 Operation 5 1 Preparations before start 5 1 1 Use the strip holders ...

Page 59: ... side down in the appropriate volume of rehydration solution with or without sample using the IPGbox Rehydration in the Cup Loading Manifold is not recommended the channel is too wide to ensure proper rehydration Also follow the instructions included with the Immobiline DryStrips or the IPGbox Close the lid of the IPGbox and allow the strips to rehydrate overnight 10 to 20 hours Ettan IPGphor 3 Op...

Page 60: ...ion of the Ettan IPGphor 3 platform near the lid hinge 2 Ensure that the manifold is level by placing the round spirit level on the center of the manifold tray after it is placed on the Ettan IPGphor 3 3 Measure out 108 ml of Immobiline DryStrip Cover Fluid even if fewer than 12 strips will be loaded into the manifold Add the cover fluid evenly in the 12 manifold channels 4 60 Ettan IPGphor 3 Oper...

Page 61: ... pointed end of the IPG strip pointing at the anode of Ettan IPGphor 3 2 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 See also the Ettan IPGphor Cup Loading Manifold User Manual 3 Ettan IPGphor 3 Operating Instructions 28 9635 51 AC 61 5 Operation 5 1 Prepara...

Page 62: ...eet over a center protrusion 1 Ensure that the feet of the cups are properly seated at the bottom of the channel Use the insertion tool wiggle the tool gently while pushing it down 2 Fill the cups with cover fluid to test for proper seating of the cups 3 Replace the cover fluid after 10 minutes 4 62 Ettan IPGphor 3 Operating Instructions 28 9635 51 AC 5 Operation 5 1 Preparations before start 5 1 ...

Page 63: ...0 μl distilled water to each paper wick 1 Position the wicks on each end of the IPG strips so that one end of the wick overlaps the end of the gel on the IPG strip 2 Ettan IPGphor 3 Operating Instructions 28 9635 51 AC 63 5 Operation 5 1 Preparations before start 5 1 2 Use the Ettan IPGphor Cup Loading Manifold ...

Page 64: ...n top of all the wicks The electrode must be in contact with the wick 1 Swivel the cams into the closed position under the external lip of the tray The electrodes should not be moved while the cams are in the closed position 2 64 Ettan IPGphor 3 Operating Instructions 28 9635 51 AC 5 Operation 5 1 Preparations before start 5 1 2 Use the Ettan IPGphor Cup Loading Manifold ...

Page 65: ...tom of the cup and come into contact with the IPG strip 2 Close the Ettan IPGphor 3 lid 3 4 Program and run the Ettan IPGphor 3 from its own control panel display See Section 5 3 Perform a run from the control panel on page68 for more details or Start the Ettan IPGphor 3 from the Ettan IPGphor 3 Control Software See Section 5 4 1 Setting up a run on page 91 for more details Ettan IPGphor 3 Operati...

Page 66: ...3 Isoelectric Focusing System is restarted Turn on power and run diagnostic program Action Step Turn on the mains Power switch located on the rear panel 1 This activates a self diagnostic program that runs for approximately 10 seconds A series of screens indicate progress If any component fails the diagnostic program will stop and a message will indicate the source of the fault 2 Note the fault if...

Page 67: ...screen in dicates that the instrument is ready as shown in the figure 5 Pr o t 1 F i l e 1 Re h y d r a t e 0 0 0 H r 2 0 C I E F 2 0 C 5 0 µ A S t r i p 1 S t r i p s 0 S t e p s Ettan IPGphor 3 Operating Instructions 28 9635 51 AC 67 5 Operation 5 2 Start up the instrument ...

Page 68: ...nel See page Section 69 5 3 1 Ettan IPGphor 3 control panel functionality 73 5 3 2 Protocol parameters 75 5 3 3 Set protocol parameters 83 5 3 4 Isoelectric focusing from the control panel 89 5 3 5 Pause or stop a protocol 68 Ettan IPGphor 3 Operating Instructions 28 9635 51 AC 5 Operation 5 3 Perform a run from the control panel ...

Page 69: ...IT The EDIT key switches the display to edit mode in which protocol steps can be programmed Refer to Section 5 3 3 Set protocol pa rameters on page 75 The EDIT key also allows the estimated run time remaining to be viewed while the unit is operating The Left arrow key moves the cursor to the left Moving the cursor off the screen to the left repositions the cursor at the left side of the next row F...

Page 70: ...display to the next group of three protocol steps Down arrow Note During electrophoresis the Up Down arrow keys are used to toggle between VhS VhT and Hrs The START button begins the selected protocol START The STOP button pauses the protocol Pressing the STOP key a second time cancels the protocol in progress STOP Press the STOP button a third time to reset the instrument to start another protoco...

Page 71: ...he LCD screen indicates the parameters of the step currently being carried out The protocol identification screen comprises the following programmable items Protocol number and name Rehydration time temperature IEF run conditions temperature and maximum current strip Number of strips and number of steps in the selected protocol P r o t 1 F i l e 1 R e h y d r a t e 0 0 0 H r 2 0 C I E F 2 0 C 5 0 ...

Page 72: ...rly with respect to time from the value set for the previous step to the value set for this step If this is the first step the value increases from 0 The original factory programmed voltage for each step is 0 Program each step with the protocol voltage values If fewer than nine steps are required end the program by as signing a value of 0 V after the final step The step duration 0 to 99 59 hours o...

Page 73: ...ommendations and protocol guidelines The permitted range of values for each pro grammable parameter is listed below Permitted range of values for each programmable parameter Recommended values Operational Range Parameter 1 to 9 Steps 10 00 to14 00 hrs Rehydration 0 to 99 59 hrs Step duration 2 00 to 10 00 hrs Focusing 0 to 99 59 hrs 100 to 10000 V 0 to 10000 V Volts 50 75 μA per strip 1500 μA tota...

Page 74: ... value from 1000 to 990 for instance even though only the ten s place is adjusted manually three digits are affected Whether individual or linked digits wrap around so increasing from 9 produces 0 and decreasing from 0 produces 9 until a parameter limit is reached Ettan IPGphor 3 can be programmed in volt hours Vhr or hours and minutes in step n hold or in gradient mode Any values within the opera...

Page 75: ...below to edit the rehydration parameters time and temperature and the IEF parameters temperature μA Strip and number of strips as necessary on the main screen Action Step Select the protocol to be created or edited from the protocol identification screen 1 Position the cursor under the protocol number field using the Right and Left arrow keys and move to the desired protocol number using the Up an...

Page 76: ...F i l e 1 R e h y d r a t e 1 0 0 0 H r 2 0 C I E F 2 0 C 5 0 µ A S t r i p 1 S t r i p s 0 S t e p s Edit time range by using the Left and Right arrow keys to position the cursor and use the Up and Down arrow keys to set the time range 4 The available time range is 0 99 59 h with 10 min resolution Hence only the first three digits are editable A minimum of 10 hrs is required for the rehydration o...

Page 77: ...form in humid environments when the platform is run below 7 C ambient temperature The third line contains fields to set the platform temperature and current limit for all isoelectric focusing steps 6 Adjust the rehydration temperature by using the Left and Right arrow keys to position the cursor Use the Up and Down arrow keys to select the value within the range of 15 C to 30 C To set the platform...

Page 78: ...rip P r o t 1 F i l e 1 R e h y d r a t e 1 0 0 0 H r 2 0 C I E F 2 0 C 5 0 µ A S t r i p 1 S t r i p s 0 S t e p s Note The total deliverable ampeerage is 1500 μA Use the arrow keys to adjust the numbers of strips to run 8 P r o t 1 F i l e 1 R e h y d r a t e 1 0 0 0 H r 2 0 C I E F 2 0 C 5 0 µ A S t r i p 1 S t r i p s 0 S t e p s Press EDIT on the control panel to move to next screen 9 78 Etta...

Page 79: ...us step group press the Up arrow key 3 To toggle to Gradient move the cursor to the Step field using the Right and Left arrow keys and toggle with the Up or Down arrow key 4 P r o t 1 3 S t e p s S 1 G r a d 5 0 0 V 1 0 0 H r s S 2 G r a d 1 0 0 0 V 4 0 0 H r s S 3 S t e p 8 0 0 0 V 8 0 0 0 0 V h r IEF voltage is programmable from 0 8000 V in increments of 10 V Each of the first three digits in th...

Page 80: ...e same manner as the Hrs values Note When toggling from Hrs to Vhr the value in the Vhrs field is automatically calculated according to the selected voltage change pattern For Step n hold Vhr is the product of hours and voltage For Gradient Vhr is the product of hours and the mean of the voltage limit set for the current step and the previous step Absolute Vhr limits can be set to any value from 1...

Page 81: ...der certain power conditions it may not be reached at all Note A lower voltage step may be entered at the end of the Ettan IPGphor 3 protocol Band sharpness will decrease however Verify the protocol Action Step Scroll through each step position the cursor in the step number field using the Right and Left arrow keys 1 Advance through the steps using the Up and Down arrow keys 2 Edit as required by ...

Page 82: ...the value in the Hrs field is 00 00 4 Exit edit mode Press EDIT to save any changes and exit the step edit mode The protocol identification screen will appear 82 Ettan IPGphor 3 Operating Instructions 28 9635 51 AC 5 Operation 5 3 Perform a run from the control panel 5 3 3 Set protocol parameters ...

Page 83: ...elect protocol number Action Step Place the cursor in the protocol number field using the Right and Left arrow keys and select the protocol number using the Up and Down arrow keys 1 P r o t 1 M Y P R O TO C O L R e h y d r a t e 1 0 0 0 H r 2 0 C I E F 2 0 C 2 0 0 µ A S t r i p 1 2 S t r i p s 3 S t e p s Use the Right and Left arrow keys to position the cursor to set the number of strips to run a...

Page 84: ... the following message will indicate the required action 5 L i d o p e n s t e p 1 C l o s e l i d t o c o n t i n u e CAUTION During IEF Do not lean on the safety lid Do not apply excess pressure or uneven weight to the lid Do not place any items on the lid Press START again after securing the safety lid 6 84 Ettan IPGphor 3 Operating Instructions 28 9635 51 AC 5 Operation 5 3 Perform a run from ...

Page 85: ... Pressing EDIT again will access the step setting screen Pressing EDIT again will return to the running screen 1 The first line indicates that the protocol is running 2 Any changes made while a protocol is running are applied to the current run only the stored protocol is not affected Move the cursor under the digit you wish to set and change it using the Up and Down arrow keys To end this step se...

Page 86: ... strip stop the run and reduce the V or μA settings The current platform temperature is indicated The average current per IPG strip μA the total instantaneous power deliv ered W and actual voltage V are indicated If the current is the limiting parameter the A in μA will flash To toggle between VhT VhS and Hrs 4 Move the cursor to the Hrs Vhrs field of the display using the Right or Left arrow key ...

Page 87: ... 0 0 H r s S 2 G r a d 2 0 0 0 V 1 0 0 H r s S 3 S t e p 8 0 0 0 V 2 0 0 H r s Note If you adjust the voltage during a gradient step the step will be converted to Step for the remainder of the step As isoelectric focusing proceeds the bromophenol blue tracking dye will migrate toward the anode and leave the IPG strip This generally occurs well before focusing is complete Note A colorless strip is ...

Page 88: ... 2 0 0 0 V h r s To t a l 5 0 µ A 8 0 0 H r To t a l P r e s s S t o p t o r e s e t The final instrument values are displayed Press STOP as indicated to reset the instrument 88 Ettan IPGphor 3 Operating Instructions 28 9635 51 AC 5 Operation 5 3 Perform a run from the control panel 5 3 4 Isoelectric focusing from the control panel ...

Page 89: ...t i n u e To continue the protocol press the START key To cancel the protocol press the STOP key again This will cause the instrument to respond as though the end of an uninterrupted protocol had been reached as described above Ettan IPGphor 3 Operating Instructions 28 9635 51 AC 89 5 Operation 5 3 Perform a run from the control panel 5 3 5 Pause or stop a protocol ...

Page 90: ...s section This section covers how you run ISO electric focusing from a PC See page Section 91 5 4 1 Setting up a run 97 5 4 2 Perform a run 90 Ettan IPGphor 3 Operating Instructions 28 9635 51 AC 5 Operation 5 4 Run a pre programmed protocol from the PC ...

Page 91: ...ndicator on the front panel should not light Main screen and buttons 4 5 1 2 3 Function Part Instrument selection buttons 1 Protocol mode buttons 2 Instrument control buttons 3 Info button 4 Ettan IPGphor 3 Operating Instructions 28 9635 51 AC 91 5 Operation 5 4 Run a pre programmed protocol from the PC 5 4 1 Setting up a run ...

Page 92: ...be shown below the buttons Up to four instruments can be run simultaneously with the software Select menu Communication Instrument 1 4 to open the instrument communication settings 3 Check Active 4 Select COM port in the COM port list box 5 Enter instrument name in the Name field 6 Click Done 7 A green lamp is lit in the corresponding instrument selection button 92 Ettan IPGphor 3 Operating Instru...

Page 93: ...e this protocol can be edited and saved to a file Use the fast mode protocol Action Step In Ettan IPGphor main window click on the appropriate instrument selection button Instr1 Instr4 to select the desired instrument 1 Select Fast Fast should be shown below the buttons 2 Select the Run settings details tab by clicking the tab 3 Set experimental conditions that is Strip length pH range and Number ...

Page 94: ...clicking the right of the two buttons at the bottom right hand corner of the tab window The protocol selected by the software is now shown in detail It is now possible to edit any values in the protocol by clicking an editable field and enter desired changes 94 Ettan IPGphor 3 Operating Instructions 28 9635 51 AC 5 Operation 5 4 Run a pre programmed protocol from the PC 5 4 1 Setting up a run ...

Page 95: ...ocol will be overwritten it will be lost Download protocols to a specific position Ettan IPGphor 3 can store up to ten protocols Action Step To download a protocol to the Ettan IPGphor 3 at another position than po sition 1 select menu Protocol Transfer protocol and select desired Instru ment position 1 10 1 Click OK 2 The current protocol is now downloaded to the selected position in Ettan IPGpho...

Page 96: ...ht button of the three buttons in the top of the Protocol view The current protocol is now deleted 96 Ettan IPGphor 3 Operating Instructions 28 9635 51 AC 5 Operation 5 4 Run a pre programmed protocol from the PC 5 4 1 Setting up a run ...

Page 97: ... downloaded to position 1 in the selected instru ment The run is now started which is indicated by the word Running below the instrument control buttons To start a second run select the desired instrument protocol and number of strips to run and click Start button It is possible to start up to four different runs using four different protocols on four different instruments in parallel 5 Ettan IPGp...

Page 98: ... is displayed below the instrument control buttons 1 The run is now stopped which is indicted by the word Idle below the instru ment control buttons 2 While using Stop it is not possible to start the protocol again except from the beginning of the protocol Note Display graphics and data Ettan IPGphor 3 Control Software provides graphics of the data Voltage Volt hours and Current as it is retrieved...

Page 99: ...tings details tab by clicking the tab heading 2 In the Protocol details graph two of three possible parameters setpoint voltage measured voltage and measured current are displayed to give an overview of the protocol and the run 3 Open any of the combo boxes below the graph and then click on the desired parameter to be displayed in the Protocol details graph Note If measured voltage and measured cu...

Page 100: ...h of the current step Open the Session log tab by clicking the tab heading to view the running data collected in the log file 6 The session log file stores instrument identity User ID current protocol start time and many other parameters please refer to Ettan IPGphor 3 Control Software User Manual for details 100 Ettan IPGphor 3 Operating Instructions 28 9635 51 AC 5 Operation 5 4 Run a pre progra...

Page 101: ...in numerical displays Store data All run data and the session log can be exported to a Microsoft Excel file It is also possible to save the current application window as an image file Follow the instructions below to export data Action Step Select Export File 1 Select file The default file name is the date and time e g Apr 11 2013 10 56 45 2 Select location The default location for the saved file ...

Page 102: ...ectrical circuits which can give a lethal electric shock Service and planned maintenance should be performed by personnel au thorized by GE WARNING Disconnect power Always disconnect power from the instrument before performing any maintenance task WARNING Do not remove the main cover There are no userserviceable components inside and you can be exposed to high voltage NOTICE Keep the instrument dr...

Page 103: ...ed Clean all other components that come in contact with the IPG strip or the sample with a detergent designed for glassware Rinse well with distilled water Clean Ettan IPGphor 3 The parts of the instrument that come in contact with liquid reagents are resistant to chemicals typically used for IEF Before introducing any other chemicals into the system first test the affected parts Cleaning procedur...

Page 104: ...is first soaked a few hours or overnight in a solution of 2 to 5 IPGphor Strip Holder Cleaning Solution in water Strip holders and manifold may also be immersed in boiling 1 SDS NOTICE You must use a neutral pH detergent such as the IPGphor Strip Holder Cleaning Solution to remove residual protein from the strip holders manifold NOTICE Fixed length strip holders and manifold may be baked boiled or...

Page 105: ...ng the electrode surfaces Wash the electrode assemblies with the IPGphor Strip Holder Cleaning Solution Rinse thoroughly with water then deionized water and allow to air dry Cleaning before planned maintenance service To ensure the protection and safety of service personnel all equipment and work areas must be clean and free of any hazardous contaminants before a Service Engineer starts maintenanc...

Page 106: ...m Section 8 1 Health and Safety Declaration Form on page112 or print it from the PDF file available on the User Documentation CD 106 Ettan IPGphor 3 Operating Instructions 28 9635 51 AC 6 Maintenance 6 1 Cleaning Ettan IPGphor 3 ...

Page 107: ...it design so it is imperative that fuses are replaced only with fuses of identical rating The mains power module located at the back of the instrument contains two input mains fuses which are the same for all models T2 5 AL 250V 5 20 mm Fuse compartment 3 1 2 4 Function Part Mains power switch 1 Hinged cover 2 Slot 3 Casette end 4 Ettan IPGphor 3 Operating Instructions 28 9635 51 AC 107 6 Maintena...

Page 108: ...ppears to be in tact check it with a multi meter A reading of 1 ohm or less indicates the fuse is still usable 3 After placing a new fuse into the cassette slide it into the power module making sure the arrow on the cassette points to the right in the same direc tion as the guide arrows on the inside of the compartment door 4 Repeat steps 2 to 4 for the second cassette 5 Close the fuse compartment...

Page 109: ...reset the instru ment Locked screen in edit mode If no electrical components are functioning e g HV ON lamp does not light and the cooling fans are motionless check the fuses in the mains power module Blank display Note the component that failed and press the START key to continue through the diagnostic program Call your local GE service for further in formation on how to remedy the failure Diagno...

Page 110: ... to remove ionic breakdown products of urea of other additives Voltage limit not reached Desalt the sample or prepare the sample so that the salt concentration is less than 10 mM Reduce the current limit Do not exceed 50 μA per strip Sparks or burning in strips Prevent the IPG strip from drying out by always applying Immobiline DryStrip Cover Fluid during rehydration Ensure that the IPG strip is f...

Page 111: ...plete technical support for all our products If you have any questions about how to use this product or would like to arrange to repair it please call or fax your local GE representative Ettan IPGphor 3 Operating Instructions 28 9635 51 AC 111 8 Reference information ...

Page 112: ...d en removed fr above has bee onnel and our c ce Engineer st e Service Engi may lead to de swer Yes or ers in box belo ous substance scanner surfac ument is clean ed to allow saf re customer to have been rem e instrument from the area Seria en cleaned to Comp Date GE and GE GE Healthc NJ 08855 1 2010 14 On Site Safety customers all arts a repair T neer upon arr lays in servicin No ow es ces or oth...

Page 113: ...ase specify service retu e system equ Wate Ethan None Argo Liqui lease d above has b ervice numbe er service Healt for Pr and or Service Ticket nel our custom aminants befo with your retu cing or return return to GE m d additional cl n in contact wi ces please sp y rn Please pro uipment er nol e empty n Helium Nit d Nitrogen Seria been cleaned t Comp instit Date er GE and GE GE Healthc NJ 08855 1 ...

Page 114: ...High voltage Up to 1 5 mA Current Working surface Specification Parameter Gold plated copper Electrode areas Up to 12 fixed length strip holders or one manifold Capacity 15 C to 30 C 2 C Platform temperature User interface Specification Item Parameter 7 key membrane keypad Control panel 4 lines 24 characters per line Liquid crystal display LCD 114 Ettan IPGphor 3 Operating Instructions 28 9635 51 ...

Page 115: ...p duration Programmable parameters 10 protocols with up to 9 steps each Protocol capacity RS232 1200 or 9600 baud 8 data bits 1 stop bit no parity no flow control 9600 baud is mandatory for connection with Ettan IPGphor 3 Control Software Serial port Ettan IPGphor 3 Operating Instructions 28 9635 51 AC 115 8 Reference information 8 2 Technical specifications ...

Page 116: ...6x 80 6499 33 Paper bridge pads pack of 20 6x 80 6498 76 Electrode set 80 6505 98 Cleaning brush 80 6194 19 Spirit level 80 6506 17 Forceps SS Accessories Code number Quantity Accessory part 80 6371 84 1 Immobiline DryStrip Reswelling Tray 7 18 cm 80 6465 32 1 Immobiline DryStrip Reswelling Tray 7 24 cm 17 1335 01 1 liter Immobiline DryStrip Cover Fluid 29 0119 27 1 liter IPGphorStripHolderCleanin...

Page 117: ...g information and for information about spare parts and accessories visit www gelifesciences com 2DE or contact your local GE representative Ettan IPGphor 3 Operating Instructions 28 9635 51 AC 117 8 Reference information 8 3 Ordering information ...

Page 118: ...holder 31 Fuses Replace 108 G General Precautions 14 Gradient 72 I IEF programming screen 72 Immobiline DryStrip Cover fluid 52 Rehydrate 59 Immobiline DryStrip Rehydrate 49 Instrument 28 International Standard 10 IPGbox 59 IPG strip manifold 35 IPG strips 51 Position 61 L LCD screen 30 67 Lid adapter 33 Light manifold 36 M Main screen Software 91 Manifold 35 60 Ceramic 35 Light 36 Manifold cerami...

Page 119: ...eneral precautions 14 Sample 65 Sample cups 62 Serial port 42 Session log 100 Software 34 Capture run data 101 Edit protocol 94 Monitor a run 99 Pause a run 98 Save protocol 95 Select instrument 92 Set up run 93 Start run 97 Stop a run 98 Store data 101 Spare parts 44 Specifications 38 Step 72 Strip holder 48 Preparation 48 Strip holders Position 55 T Transport 40 Typographical conventions 6 U Unp...

Page 120: ...st Contact your local GE Healthcare repre sentative for the most current information GE Healthcare Bio Sciences AB Björkgatan 30 751 84 Uppsala Sweden GE Healthcare Europe GmbH Munzinger Strasse 5 D 79111 Freiburg Germany GE Healthcare Bio Sciences Corp 800 Centennial Avenue P O Box 1327 Piscataway NJ 08855 1327 USA GE Healthcare Japan Corporation Sanken Bldg 3 25 1 Hyakunincho Shinjuku ku Tokyo 1...

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