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Ettan IPGphor 3 Isoelectric Focusing System function and description

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Ettan IPGphor 3 Instrument User Manual 11-0034-58 Edition AB

11

Fig 1-4.

Components of the Ettan IPGphor Cup Loading Manifold system. Only one of each

item shown.

A.

B.

C.

E

F

H.

I.

D.

G

J

A. Manifold, ceramic or light

F. Pre-cut paper bridges

B. Pre-cut electrode wicks

G. Sample cup insertion tool

C. Electrode assembly

H. Cleaning solution

D. Sample cups

I. Forceps

E. Cleaning brush

J. Spirit level

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Содержание Ettan IPGphor 3

Страница 1: ...GE Healthcare Ettan IPGphor 3 Instrument User Manual Ettan...

Страница 2: ......

Страница 3: ...ment It is important not to continue until all stated conditions are met and clearly understood Note The Note sign is used to indicate information important for trouble free and optimal use of the pro...

Страница 4: ......

Страница 5: ...ctrode paper wicks 24 3 2 6 Positioning the electrodes 24 3 2 7 Loading samples into the sample cups 24 3 3 Using fixed length Ettan IPGphor 3 strip holders 25 3 3 1 Preparing the strip holder s 25 3...

Страница 6: ...enance 4 1 Cleaning the Ettan IPGphor 3 49 4 2 Cleaning procedures 50 4 2 1 Ceramic strip holders manifold 50 4 2 2 Manifold Light 51 4 2 3 Electrodes 51 4 3 Replacing fuses 52 5 Troubleshooting 6 Cus...

Страница 7: ...oteins and other light sensitive stains IEF runs are programmed from the Ettan IPGphor 3 control panel with up to ten user defined IEF protocols each with up to 9 steps per protocol programmable funct...

Страница 8: ...ed the log files are exported to programs such as Microsoft Excel The Ettan IPGphor 3 Control Software controls up to four Ettan IPGphor 3 units at one time each running a different set of run paramet...

Страница 9: ...tents with the packing list If any part is missing please contact your local GE Healthcare sales office Inspect all components If any part appears damaged please contact the courier immediately Please...

Страница 10: ...n 10 Ettan IPGphor 3 Instrument User Manual 11 0034 58 Edition AB Fig 1 2 Light protective cover Fig 1 3 Location of mains power switch and serial port Light protective cover L The mains power switch...

Страница 11: ...n AB 11 Fig 1 4 Components of the Ettan IPGphor Cup Loading Manifold system Only one of each item shown A B C E F H I D G J A Manifold ceramic or light F Pre cut paper bridges B Pre cut electrode wick...

Страница 12: ...2 Ettan IPGphor 3 Instrument User Manual 11 0034 58 Edition AB Fig 1 5 Strip holders Note All strips and holders used at any one time must be of the same length Fig 1 6 Lid adapter Five strip holder l...

Страница 13: ...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 On the bo...

Страница 14: ...the correct amount of pressure to the lids of the Ettan IPGphor 3 strip holders Ettan IPGphor 3 Control Software Ettan IPGphor 3 Control Software allows the user to control the Ettan IPGphor 3 Isoele...

Страница 15: ...ample cup accommodates sample volumes of up to 150 l IEF electrode paper wicks The small precut rectangular wicks Fig 1 4 must be placed at both the anodal and cathodal ends of the rehydrated IPG stri...

Страница 16: ...tion volume i e 30 l maximum for both wells Introduce the sample through the wells if it is not included in the rehydration solution Cover The cover is made of acrylic and protects the IPG strip durin...

Страница 17: ...radient and step duration Protocol capacity 10 protocols with up to 9 steps each Serial port RS232 1200 or 9600 baud 8 data bits 1 stop bit no parity no flow control 9600 baud is mandatory for connect...

Страница 18: ...nit from a PC The Ettan IPGphor Cup Loading Manifold User Manual contains instructions for first dimension isoelectric focusing of proteins on IPG strips The Ettan IPGphor 3 Safety Handbook contains s...

Страница 19: ...er outlets 1 Select the cord with the appropriate plug for the power outlet in your laboratory If neither of these cords are suitable for your power outlets obtain a detachable cord with an IEC 320 C1...

Страница 20: ...required For any other configuration a null modem cable is required Pin number assignments and device set up details follow To set the baud rate 1 Press the Up and Down arrows together to access the...

Страница 21: ...he source of the fault note the fault and press the START key to skip to the next test If any failure is detected call GE Healthcare service 3 1 1 Ettan IPGphor 3 control panel Once the diagnostic pro...

Страница 22: ...first protocol steps The EDIT key also allows the estimated run time remaining to be viewed while the unit is operating The up arrow key increases the value of the selected field or digit and the dow...

Страница 23: ...the Ettan IPGphor 3 platform The small T shaped protrusion fits into a cutout section of the Ettan IPGphor 3 platform near the lid hinge 3 Ensure that the manifold is level by placing the round spirit...

Страница 24: ...overlaps the end of the gel on the IPG strip 3 2 6 Positioning the electrodes 1 With the electrode cams in the open position position the electrode assembly on top of all the wicks The electrode must...

Страница 25: ...he strip holder s corresponding to the IPG strip length chosen for the experiment 7 11 13 18 or 24 cm 2 Wash each holder with the strip holder cleaning solution supplied to remove residual protein 3 R...

Страница 26: ...attached to each Immobiline DryStrip package 1 Deliver the solution slowly at a central point in the strip holder channel away from the sample application wells 2 Remove any larger air bubbles Positio...

Страница 27: ...der Pressure blocks on the underside of the cover ensure that the strip maintains good contact with the electrodes as the gel rehydrates Rehydration proceeding Rehydration can proceed on the bench top...

Страница 28: ...ak with deionized water Remove excess water by blotting with tissue paper 3 Using forceps or tweezers lift one end of the rehydrated IPG strip Position an electrode pad over the electrode then lower t...

Страница 29: ...rip holder on the platform as shown below The pointed end of the holder 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 platf...

Страница 30: ...er To ensure good electrical contact when using the fixed length strip holder 1 Apply two lid adapters right across the lids of the strip holders one over the anodic area and the other over the cathod...

Страница 31: ...4 58 Edition AB 31 To inspect the run the light protective cover see figure below can be lifted without stopping the run To open the safety lid Press down the lid and the lock is released and the run...

Страница 32: ...adient Editing of the protocols and transferring them to the instrument Starting stopping and pausing of IEF runs on the instrument Acquisition of run data which is displayed in the graph and the nume...

Страница 33: ...rom the menu Select an instrument Select the desired Ettan IPGphor 3 instrument by clicking one of the instrument selection buttons The identity of the instrument should now be shown below the buttons...

Страница 34: ...t the optimal protocol for these parameters Edit selected protocol Select the Protocol view in the Run settings details tab by clicking the right of the two buttons at the bottom right hand corner of...

Страница 35: ...red voltage and measured current can be displayed by selecting from the combo boxes below the graph Ettan IPGphor 3 Control Software graphs the data Voltage Volt hours and Current as it is retrieved f...

Страница 36: ...and the data is presented in a graph and in numerical displays Storing data All run data and the session log can be exported to a Microsoft Excel file It is also possible to save the current applicati...

Страница 37: ...ually adjustable digits or characters Certain digits are also linked to their neighbouring digits For example three of the four digits in the voltage field can each be adjusted individually but when a...

Страница 38: ...t the characters as desired To skip editing the name use the left arrow to proceed to the next line 4 To change any letter move the cursor to that location using the right and left arrow keys and then...

Страница 39: ...ation to 00 00 Hrs To adjust the rehydration temperature 1 use the left and right arrow keys to position the cursor and 2 the up and down arrow keys to select the value within the range of 15 30 C The...

Страница 40: ...the numbers of strips to run 4 To advance to the next edit screen press the EDIT key IEF programming The second screen in edit mode is the first in a series of nine programmable IEF steps in which the...

Страница 41: ...st move the cursor to the desired digit location using the right and left arrow keys and 2 then change the value using the up and down arrow keys The original factory programmed voltage for each step...

Страница 42: ...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 30000 Vhr Programming example Simplified 4 step pr...

Страница 43: ...verify the protocol 1 scroll through each step position the cursor in the step number field using the right and left arrow keys and 2 then advance through the steps using the up and down arrow keys Ed...

Страница 44: ...he up and down arrow keys to select the number 1 12 4 Press the START key A screen is displayed to confirm the number of strips to run Adjust the number of strips if necessary 1 Press START to continu...

Страница 45: ...s 1 Press the EDIT key to display the set screen The first line indicates that the protocol is running Pressing Edit again will access the step setting screen Pressing Edit again will return to the Ru...

Страница 46: ...ll strips and will often read 0 W because any value less than 0 5 is rounded down to 0 If the delivered power exceeds 0 4 W per strip stop the run and reduce the V or A settings The current platform t...

Страница 47: ...ring 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...

Страница 48: ...Ettan IPGphor 3 Control Software An abbreviated log illustrates the format Ettan IPGphor 3 Control Software may also be used to log and graph and print the output In addition the output may be export...

Страница 49: ...RTANT The ceramic strip holders and manifold are very brittle and fragile Take care in handling and do not subject to impacts IMPORTANT Avoid scratching the electrode surfaces IMPORTANT Wipe the exter...

Страница 50: ...ld is first soaked a few hours or overnight in a solution of 2 5 IPGphor Strip Holder Cleaning Solution in water Strip holders and manifold may also be immersed in boiling 1 SDS Cleaning procedure 1 F...

Страница 51: ...ng Solution 4 2 3 Electrodes Wash the electrode assemblies with the IPGphor Strip Holder Cleaning Solution Rinse thoroughly with water then deionized water and allow to air dry CAUTION Do not use any...

Страница 52: ...heck it with a multi meter A reading of 1 ohm or less indicates the fuse is still usable 4 After placing a good fuse into the cassette slide it into the power module making sure the arrow on the casse...

Страница 53: ...r the second cassette 6 Close the fuse compartment cover and gently press it into the power module until it snaps shut 7 Plug the power cord into the unit and turn the mains power switch on 8 If the i...

Страница 54: ...4 Care and maintenance 54 Ettan IPGphor 3 Instrument User Manual 11 0034 58 Edition AB...

Страница 55: ...been detected The instrument will reduce the voltage setting 500 volts automatically The run continues and there is no need for further action Power delivery Current too low or zero At least two of t...

Страница 56: ...he entire length of the strip The IPG strip should be in complete contact with the correct volume of rehydration solution Remove any air bubbles trapped under the IPG strip Desalt the sample or prepar...

Страница 57: ...s properly completed Product Quantity Code number Ettan IPGphor 3 Isoelectric Focusing Unit Basic unit Order manifold and strip holders separately 1 11 0033 64 Cable Serial 9 Pin F F with Null Modem A...

Страница 58: ...9 88 7 cm complete 1 80 6416 87 11 cm complete 1 80 6417 06 13 cm complete 1 80 6417 25 18 cm complete 1 80 6417 44 24 cm complete 1 80 6470 07 Immobiline DryStrip Reswelling Trays 7 18 cm 1 80 6371 8...

Страница 59: ...483 56 Mini Dialysis Kit 1 kDa cut off 250 l 1 80 6483 75 Mini Dialysis Kit 1 kDa cut off 2 ml 1 80 6483 94 Mini Dialysis Kit 8 kDa cut off 250 l 1 80 6484 13 Mini Dialysis Kit 8 kDa cut off 2 ml 1 80...

Страница 60: ...12 17 6001 15 pH 3 11 NL 12 17 6003 75 pH 4 7 12 17 6001 13 pH 5 3 6 5 12 17 6003 60 pH 6 11 12 17 6001 96 pH 6 2 7 5 12 17 6003 65 pH 7 11 NL 12 17 6003 70 18 cm pH 3 5 6 12 17 6003 56 pH 3 10 12 17...

Страница 61: ...4 7 12 17 6002 46 pH 4 5 5 5 12 17 6002 40 pH 5 0 6 0 12 17 6002 41 pH 5 3 6 5 12 17 6003 62 pH 5 5 6 7 12 17 6002 42 pH 6 9 12 17 6002 47 pH 6 2 7 5 12 17 6003 67 pH 7 11 NL 12 17 6003 72 Cleaning so...

Страница 62: ...6 Customer service information 62 Ettan IPGphor 3 Instrument User Manual 11 0034 58 Edition AB...

Страница 63: ......

Страница 64: ...away NJ 08855 1327 USA GE Healthcare Sanken Bldg 3 25 1 Hyakunincho Shinjuku ku Tokyo 169 0073 Japan Instrument User Manual 11 0034 58 AB Asia Paci c Tel 852 2811 8693 Fax 852 2811 5251 Australasia Te...

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