Hoefer IEF100 Manuel d'utilisateur

Hoefer IEF100 Isoelectric Focusing Unituser manualum IEF100-IM/Rev.B0/07-12

• p4Level230 V Power Cords (EU and UK)Large Rehydration/Equilibration Tray IEF110Electrode Wicks IEF106Sample Cups IEF108ForcepsBrush(+) ElectrodeAcce

• p5Component DescriptionsSafety LidThe transparent safety lid must be closed in order for voltage to be applied to the electrodes. Interlocks are use

• p6STOP ButtonImmediately stops the IEF, and terminates the protocol.LCD DisplayThe large blue/white display simplifies the interface, and displays gr

• p7Focusing Tray The 6 channel focusing tray accommodates IPG strips up to 24 cm in length. Small grooves in the tray allow for easier removal of the

• p8ElectrodesPositive Electrode (Anode)A high volt connector attaches the positive (+) electrode(s) to the high volt terminal. The anode has a single

• p9Electrode WicksThe electrode wicks are small rectangular filter papers that are placed over the end of the IPG strips. The electrodes make contact

• p10Programming the IEF100Main ScreenWhen turned ON, the main screen is displayed (Fig. 10). The main screen contains the following fields. Date and T

• p11The Four Buttons: EDIT, RUN, OPTION and GRAPHEDITEdits the highlighted protocol.RUNStarts the highlighted protocol.OPTIONAllows access to port se

• p12Edit Protocol ScreenFrom the main menu, use the knob to highlight the desired protocol. Press the button beneath EDIT on the screen. The IEF100 w

• p13The lower case “m”, “u”, and “n” can be used to designate milli, micro, and nano, if desired. When the name is correct, click the knob to enter t

• piPage Finder Important Information ...iiWaste Electrical And Electronic Equipment (WEE

• p14StepsTo edit any step parameter, turn the knob to highlight the parameter. Press the knob, and turn the knob to set the desired value. Press the

• p15Step Time Steps can be programmed to run for a set amount of time, Hrs (00:00 in hours:minutes), or for a set amount of volt-hours, Vhrs (volts m

• p16Run ScreenThe information on the run screen is described in detail in the IEF operation section. The run screen has no editable fields. It does ha

• p17Options ScreenFrom the main menu, pressing the OPTION key will allow access to the following options (Fig. 13).Serial Port and IntvlEthernetDate/

• p18To Set the DateThe date field is three separate fields of DAY/MONTH/YEAR. 1 Use the knob to highlight the date field. Click the knob to set the da

• p19Clear All ProtocolsThis option is used to overwrite the existing protocols in memory. A popup screen shows three options:Defaults Erase Cancel• H

• p20IEF100 OperationUsing the KnobThe knob has two actions, it turns, and pushes in to “click” (Fig. 14).Turn: Navigates through fields, adjusts value

• p21Preparing IPG StripsImmobilized pH Gels (IPG) are ultra thin polyacrylamide gels on plastic support films. The gels are made with a pH gradient co

• p22Note: Rehydration solutions typically contain high concentrations of urea that tend to crystallize at lower temperatures. For this reason, rehyd

• p23Isoelectric Focusing (using the Single Electrode Set)1Open the safety lid by pressing down on the white push label in the middle of the front of

• piiImportant Information – English• If this equipment is used in a manner not specied by Hoefer, Inc. the protection provided by the equipment may

• p245Align the (+) end of the strips with the alignment mark in the tray (Fig. 20). This will help in two ways. First, the clamping tabs will not int

• p25Note: The electrodes will not make contact with the IPG strips without the electrode wick in place.7Connect the (+) electrode to the (+) termina

• p269Connect the (–) electrode to the (–) terminal (Fig. 25).0Place the (–) electrode on top of the electrode wicks so that the platinum wire is cent

• p27Isoelectric Focusing (using the Dual Electrode Accessory)The Dual Electrode Accessory increases the Hoefer IEF100 capacity, enabling up to twelv

• p286The second set of IPG strips are loaded to the right of the first set in the opposite orientation — with the anodic end (+) of the strips matchin

• p29Sample CupsSample cups can be used to apply protein sample to IPG strips if it was not included in the rehydration buffer. The sample cups are su

• p30Start IEF1 Highlight the desired protocol, and press RUN.2 Confirm the number of focusing tray channels containing IPG strips. Use the knob to cha

• p31Run ScreenThe run screen displays all the information on the current run (Fig. 33). There are no editable fields. The information on the run scre

• p32In the example below 6 IPG strips are being focused according to protocol 7, a maximum of 0.5 W/strip and 100,000 volt-hour endpoint (Fig. 34). T

• p33Typical Isoelectric FocusingThe first step is typically set to low voltage to limit the current and let the ions move to the end of the IPG strip

• piiikäyttöjännitelyijyjä käyttöjännitteeseen. • Kiertää kaikki käyttöjännitevalvonnat ja irrottaa valtalyijyt ennen poistaminen turvallisuuskantta.

• p34When focusing is complete, the IEF100 will beep, and an end run screen will be displayed (Fig. 36). Line 1The protocol number and name. The word

• p35Optional Data ConnectionsSerial Printer ConnectionUse the RS232 port to connect directly to a serial printer. The printer and cable are not suppl

• p36Serial Port Configuration1 Select a Com Port (Fig. 38). This is the computer port to communicate directly with the IEF100. Choose TCP/IP if con

• p37TCP/IP Connection Using the Ethernet PortUse the IEF100 ethernet port to connect to a local area network (LAN). Plug a live ethernet cable into t

• p384 Click on the “SETTINGS” tab and the “ASCII Setup” button (Fig. 42).5 In the ASCII setup section, check the boxes “Send line ends with line

• p39Note: The data from the IEF100 is a series of text fields separated by commas. One method to transfer data to other programs is to use the Window

• p40HyperTerminal CommandsThe HyperTerminal program communicates with the IEF100 by trans-mitting one line of data at a time.Multiple lines can be ty

• p41Subset of HyperTerminal Commands Format of Command Command Description Expand Description Result of Command CommandAT Check connect

• p42Care and Maintenance Turn the mains power switch off and unplug the power cord before cleaning. Instrument and Lid• Use a soft cloth dampened w

• p43Troubleshooting Problem SolutionInstrument does not turn on Check power cord is plugged into grounded outlet and ON/OFF switch is turned to t

• pivtrykket er unregulated. • Introduserer Aldri antifreeze eller noe organisk løsemiddel inn i noe del av instrumentet. Organiske løsemiddler vil f

• p44Problem SolutionSet voltage not reached Continue with the IEF. IPG strip is being limited by μA or W. Conductivity of the strip is too high.

• p45Ordering Information Product Quantity Order CodeIEF100 1 IEF100Running Tray 1 IEF105Electrode Wicks 504 IEF106Sample Loading Cups (10 st

• p46Appendix A: Preprogrammed ProtocolsThe following 9 protocols are preprogrammed. They are to be used as guidelines.Each protocol has a 1 hour, 10

• p47Program 5Focusing of 18 cm IPG’S under constant watt conditions.Name: 18 cm const watt Delay 0:00, Delay temp 20 °C, Run temp 20 °C, 500 μA, 1200

• p48Pre-programmed Protocols in Machine Code Format Protocol 1IPROT,1,7cm broad&basic,D,0:00,20,20,500,6000,0.5IPROT,1,7cm broad&basic,1,G,5

• p49Protocol 4IPROT,4,18cm narrow,D,0:00,20,20,500,12000,1.5IPROT,4,18cm narrow,1,G,1000,H,1:00IPROT,4,18cm narrow,2,G,12000,H,1:00IPROT,4,18cm narro

• p50Protocol 7IPROT,7,24cm narrow,D,0:00,20,20,500,12000,2.0IPROT,7,24cm narrow,1,G,1000,H,1:00IPROT,7,24cm narrow,2,G,12000,H,1:00IPROT,7,24cm narro

• p51Appendix B: Reagents and SolutionsSample PreparationSamples prepared for 2D should be fully denatured, free of insoluble material, and low in ove

• p52General Guidelines • Sample preparation procedures are being refined and standardized. It is best to consult the literature to determine if a par

• p53RecipesIEF Sample Extraction Buffer for 2D1A. Urea Sample Buffer Solution Prepares 25 ml 9.5 M urea, 4% CHAPS Final Concentration AmountUrea (F

• pvWaste Electrical And Electronic Equipment (WEEE) This symbol indicates that the waste of electrical and electronic equipment must not be disposed

• p54IPG Strip Rehydration Solution2A. Urea Rehydration Stock Solution Prepares 50 ml 8 M urea, 2% CHAPS, 0.002% bromophenol blue Final Concentratio

• p553. SDS Equilibration Buffer SolutionThis solution is used after IEF, and before second dimension PAGE. The IPG strips are immersed in excess solu

• p56Agarose Overlay 1% Agarose in 1X Electrophoresis Buffer Prepares 100 ml 1% agarose, 25 mM Tris, 192 mM glycine, 0.1% SDS Prepare in a 500 ml flask

• p57Appendix C: References IEF100 Ames, G. F. L. and Nikaido, K. Two-dimensional gel electrophoresis of membrane proteins. Biochemistry 15, 616–623 (

• p58Görg, A., Obermaier, C., Boguth, G., Posch, A. and Weiss, W. Two-dimensional polyacrylamide gel electrophoresis with immobilized pH gradients in

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• p1Function and DescriptionThe Hoefer IEF100 Isoelectric Focusing instrument is designed to perform isoelectric focusing of proteins in Immobilized p

• p2Specifications Capacity using Single Electrode set 1– 6 IPG strips 7 to 24 cm longCapacity using Dual Electrode accessory 2– 12 IPG strips

• p3System Components Unpacking Unwrap all packages carefully and compare the contents with the packing list, making sure all items arrived. If any pa