Ettan IPGphor Cup Loading Manifold
Contents
1. Immobiline DryStrip gels Running conditions for application method Cup Loading Bridge Loading Length pH range s Voltage mode Voltage Duration Volt hours Duration Volt hours cm V h min kVh h min kVh Y 3 10 Gradien 500 0 0 0 0 0 01 0 0 3 10 NL 2 Gradien 4000 30 3 4 2 30 5 6 4 7 3 Step and Hold 5000 0 45 3 7 0 30 2 5 6 11 Tota 2 15 7 1 3 00 8 0 11 3 10 Gradien 500 0 0 0 0 0 01 0 0 4 7 2 Gradien 4000 30 3 4 2 30 5 6 3 Step and Hold 8000 30 0 6 1 40 2 0 Tota 3 00 4 0 4 10 7 6 6 11 Gradien 500 0 0 0 0 0 01 0 0 2 Gradien 4000 30 3 4 2 30 5 6 3 Step and Hold 8000 15 8 5 1 30 0 0 Tota 2 45 1 9 4 00 5 6 13 3 10 Gradien 500 0 0 0 0 0 01 0 0 3 10 NL 2 Gradien 4000 30 3 4 2 30 5 6 4 7 3 Step and Hold 8000 50 3 5 2 10 5 2 Tota 3 50 6 9 4 40 20 8 6 11 Gradien 500 0 0 0 0 0 01 0 0 2 Gradien 4000 30 3 4 2 30 5 6 3 Step and Hold 8000 40 1 6 1 50 3 4 Tota 3 10 5 0 4 20 9 0 18 3 10 Step and Hold 300 3 00 0 9 3 00 0 9 3 10 NL 2 Gradien 1000 6 00 3 9 6 00 3 9 4 7 3 Gradien 8000 3 00 3 5 3 00 3 5 4 Step and Hold 8000 1 10 9 7 1 50 4 7 Total 13 10 28 0 13 50 33 0 6 9 1 Step and Hold 300 3 00 0 9 3 00 0 9 2 Gradien 1000 6 00 3 9 6 00 3 9 3 Gradien 8000 3 00 13 5 3 00 3 5 4 Step and Hold 8000 3 20 26 7 4 00 31 7 Total 15 20 45 0 16 00 50 0 6 11 1 Step and Hold 500 1 00 0 5 3 00 1 5 2 Gradien 1000 2 00 1 5 2 00 1 5 3 Gradien 8000 3 00 13 5 3 00 13 5 4 Step and Hold 8000 1 05 8 5 1 30 11 5 T2. manual 2 D electrophoresis Ettan IPGphor Cup Loading Manifold Ettan IPGphor Isoelectric Focusing System e A Oo Lio Amersham um 80 6499 52 Rev AO 06 03 ec o IIOS Page finder Ettan IPGphor Cup Loading Manifold 1 IPGphor unit Modifications iure RR Rare 5 Instr ctions scien eRe A 6 Suggested run conditions 244 444 a AG E n 10 TROWDIESHOOUIMS 4 gt aiar che ct aay ah coin ak carat NS ma AA a eds et ds a a 11 Paper bridge loading 12 Care amd Cleaning u KAG NAAN a Naa adie aga ada ae bar 13 Customer service information 13 Ordering information ip cus 0 3 dk an rn aa a a de 13 Ettan IPGphor Cup Loading Manifold e safety warnings and precautions English Fran ais um 80 6499 52 pii Safety warnings and precautions Important user information Please read this entire manual to fully understand the safe and effective use of this product The exclamation mark within an equilateral triangle is intended to alert the user to the pres ence of important operating and maintenance instructions in the literature accompanying the instrument If you have any comments on this manual please send them to us at Amersham Biosciences AB SE 751 84 Uppsala Sweden Amersham Biosciences reserves the right to make changes in the specifications without prior notice Warranty and
3. Request a copy of the Amersham Biosciences Health and Safety Declaration form before returning the item No items can be accepted for servicing or return unless this form is properly completed e care and cleaning Care and cleaning Ceramic Manifold Gently clean the manifold after each use Wash the manifold channels with the Ettan IPGphor Strip Holder Cleaner Rinse thoroughly with water and then deionized water and allow the manifold to air dry Do not use abrasive cleaning agents or pads Take care to avoid chipping the manifold Electrodes Wash the electrode assemblies with the Ettan IPGphor Strip Holder Cleaner Rinse thoroughly with water then deionized water and allow to air dry Customer service information Technical service and repair Amersham Biosciences offers complete technical support for all our products If you have any questions about how to use this product please call or fax your local Amersham Biosciences representative Ordering information Replacement parts Code number Ettan IPGphor Cup Loading Manifold 80 6498 57 Sample cups pack of 20 6x 80 6498 95 Paper electrode wicks pack of 40 6x 80 6499 14 Paperbridge pads pack of 20 6x 80 6499 33 Electrode set 80 6498 76 Lid adapter 80 6499 71 Cleaning brush 80 6505 98 Spirit level 80 6194 19 Forceps SS 80 6506 17 Accessory products Immobiline DryStrip Reswelling Tray 7 18 cm 1 80 6371 84 Immobiline Dr
4. en el manual advierte al usuario sobre la presencia de instrucciones importantes de operaci n y mantenimiento del aparato Si desearan hacer alg n comentario sobre este manual tengan la amabilidad de remitirlo a Amersham Biosciences AB SE 751 84 Uppsala Sweden Amersham Biosciences se reserva el derecho a modificar las specificaciones sin previo aviso Garantia y responsabilidad Amersham Biosciences garantiza que el producto entregado ha sido probado a fondo para comprobar el cumplimiento de las especificaciones publicadas La garantia incluida en las condiciones de entrega s lo es v lida si el producto se ha instalado y utilizado de acuerdo con las instrucciones entregadas por Amersham Biosciences Amersham Biosciences no ser responsable bajo ning n concepto de dafios directos o indirectos incluyendo sin limitaci n la p rdida de beneficios la p rdida de ingresos la p rdida de oportunidades de negocio la p rdida de utilizaci n y otras consecuencias relacionadas cualquiera que sea la causa que se deban a la utilizaci n defectuosa e incorrecta del producto No est permitida la reproducci n ni el almacenaje en un sistema de recuperaci n ni la transmisi n de parte alguna de esta publi Wichtige Benutzerinformationen F r ein vollst ndiges Verst ndnis und eine sichere Handhabung dieses Produktes ist es notwendig da der Benutzer dieses Handbuch vollst ndig durchliest Ein Ausrufezeichen in einem gleichseitige
5. liability Amersham Biosciences guarantees that the product delivered has been thoroughly tested to ensure that it meets its published specifications The warranty included in the conditions of delivery is valid only if the product has been installed and used according to the instructions supplied by Amersham Biosciences Amersham Biosciences shall in no event be liable for incidental or consequential damages including without limitation lost profits loss of income loss of business opportunities loss of use and other related exposures however caused arising from the faulty or incorrect use of the product 2002 Amersham Biosciences Corp All rights reserved No part of this publication may be reproduced stored in a retrieval system or transmitted in any form by any means without permission in written form from the company Renseignements importants d utilization Pour une bonne compr hension et une utilisation en s curit maximale il convient de lire enti rement ce manuel Dans la documentation qui accompagne l instrument un point d ex clamation dans un triangle quilat ral a pour but d attirer l attention de l utilisateur sur des instructions importantes de fonctionnment ou de maintenance Tous vos commentaires sur ce manuel seront les bienvenus et veuillez les adresser a Amersham Biosciences AB SE 751 84 Uppsala Sweden Amersham Biosciences se r serve le droit d effectuer des modifications de ces sp cifications sa
6. positioning anodic end on the paper bridges Note A single paper bridge can be used with the 24 cm gel strip A paper bridge can be used on both ends of all other strips at one time um 80 6499 52 pl2 Paper bridge loading Large sample volumes and large protein amounts can be applied using paper bridge loading A paper pad paper bridge is soaked with sample and placed between the anodic end of the Immobiline DryStrip gel and the electrode 375 500 pl sample can be applied using the paper bridge pads supplied with the manifold Solutions containing up to 5 mg protein have been loaded on a 18 cm long narrow pH range Immobiline DryStrip gel The rehydrated Immobiline DryStrip gel is first positioned in the bottom of the manifold channel Then the paper bridge with sample is positioned followed by a paper wick Fig 16 With anodic application the anode electrode is positioned as far out as possible in the electrode assembly while the cathode electrode is positioned close to the end of the Immobiline DryStrip gel to ensure good contact between the paper wick and Immobiline DryStrip gel electrode assembly sample cup paner imm AE as anode paper bridge hold down teeth Important Do not use strong acids bases ketones alcohols or other reagents to clean the covers sample cups or electrodes or the parts may be damaged Cups may be briefly rinsed with ethanol if desired Important
7. across the bottom of the electrode teeth to provide electrical contact to the IPG strips The hold down teeth are found along the opposite length of the electrode They are used to apply pressure on the paper bridges when the manifold is used for bridge load ing Cams on the sides of the electrode secure the electrodes in place and must be in the closed position during IEF cup arms Fig 4 Sample loading cups Fig 5 Sample cup insertion tool K Fig 6 Paper wicks e Ettan IPGphor Cup Loading Manifold Sample Cups Sample cups Fig 4 can be placed almost anywhere along the length of the Ettan IPGphor manifold that is not blocked by a centering protru sion The sample cups are supplied in strips of 6 for easy handling and placement If fewer than 6 cups are required they may be easily sepa rated by cutting the thin plastic bridging the strip of cups together The sample cups can accommodate sample volumes of up to 150 pl For proper sealing of the cup to the gel all of the feet of the sample cup must rest on the bottom of the channel and all cup arms must be fully pressed down into the channel Sample cup insertion tool The sample cup insertion tool Fig 5 is supplied with the manifold kit and refill packages of sample cups It is used for proper placement and sealing of the cup over the IPG strips IEF electrode paper wicks The small precut rectangular wicks Fig 6 must be placed at both the anodal and catho
8. annel the end of the gel not the end of the plastic should align with the etched mark 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 Fig 11 Oc SA 5 N 31818 88 8 8 8 GHB 24 cm 18 cm 13 cm 11 cm Ettan IPGphor Cup Loading Manifold e instructions Fig 12 Sample cup positioning details Note Cups must not straddle the centering protrusions on the bottom of the channels um 80 6499 52 p8 Place a strip of cups in the appropriate position Fig 12 for example 1 cm from the end of the gel portion of the IPG strip Do NOT place the cup with the feet over a center protrusion Push the cups into the channels with gloved fingers starting at one end of the strip and working towards the other Align the insertion tool over the cups and push down to ensure that the feet of the cups are properly seated at the bottom of the channel wiggle the tool gently while pushing down in order to ensure that the cups are seated as far down as they will go Take care not to move the cups while removing the insertion tool Fill the cups with cover fluid to test for proper seating of the cups Remove the cover fluid after 10 min
9. d channel Check for correct positioning of sample cup arms Check that the feet of the cups are not resting on a centering protrusion in the channel Check that the strip is centered inside of the channel Sparking or burning in the Immobiline DryStrip gels Current limit setting is too high Immobiline DryStrip gel is not fully rehydrated Immobiline DryStrip gels dried out during IEF Do not exceed the maximum recommended setting of 50 pA per Immobiline DryStrip gel Ensure that the Immobiline DryStrip gels are rehydrated with a sufficient volume of rehydration solution Remove any large bubbles trapped under the Immobiline DryStrip gel after placing it on rehydration solution Always apply Immobiline DryStrip Cover Fluid to prevent dehydration of rehydrated Immobiline DryStrip gels Immobiline DryStrips turn white and opaque after focusing Immobiline DryStrip gels dried out during IEF Always apply recommended amount of Immobiline DryStrip Cover Fluid to prevent dehydration of rehydrated Immobiline DryStrip gels Immobiline DryStrip Cover Fluid overflows from Cup Loading Manifold Excess cover fluid added Do not add more than the recommended volume Ensure that the outside rim of the tray does not have any oil on it pll Ettan IPGphor Cup Loading Manifold e paper bridge loading Note The application point anodic or cathodic is an important factor for obtaining good results Fig 16 Electrode
10. dal ends of the rehydrated IPG strips just under the elec trodes The wicks absorb excess water salts and proteins with pIs that lie outside the pH range of the IPG strip p3 Ettan IPGphor Cup Loading Manifold e Ettan IPGphor Cup Loading Manifold FG Fig 7 Paper bridges Fig 8 Standard strip holder adapter um 80 6499 52 p4 Paper bridge loading pads Bridge loading pads Fig 7 are used to load larger volumes of sample 375 500 pl than what can be held in a sample cup The bridges are 0 8 cm x 5 0 cm in size with pointed ends One end of the pad contacts the IPG strip and is held in place by the hold down teeth of the electrode assembly The other end of the bridge loading pad makes contact with a paper wick placed under the platinum wire of the electrode assembly Lid adaptor for standard strip holders When using the IPGphor standard strip holders or the IPGphor cup load ing strip holders a foam pad lid adapter Fig 8 must be used to apply the correct amount of pressure to the tops of the IPGphor strip holders and keep them in contact with the IPGphor power supply pads If running only one IPGphor strip holder a second IPGphor strip holder must be placed on the opposite side of the lid adapter to maintain the adapter in a level position Not included but required Immobiline DryStrip Cover Fluid order separately Immobiline DryStrip Cover Fluid is required to ensure that the rehy drated IPG strip gels
11. do not dry out during electrophoresis Without cover fluid the strips will dry out urea will crystallize and the sample will not focus properly Immobiline DryStrip Reswelling Tray order separately The Immobiline DryStrip Reswelling Tray is required for proper strip rehydration The channel in the Cup Loading Manifold is too wide to ensure proper absorption of the required volumes of rehydration solution Two trays are available One for 7 18 cm strip lengths and a second for 7 24 cm strip lengths Fig 9 Lid modifications e IPGphor lid modifications IPGphor lid modifications The IPGphor lid must be modified before using the manifold system The three 3 foam pads on the lid must be removed Fig 9 Once removed any adhesive remaining on the lid must be cleaned off The foam pads are NOT replaced on the lid After lid modification the lid adaptor Fig 8 must be used when running IPGphor strip holders Ettan IPGphor Cup Loading Manifold e instructions Fig 10 Manifold placement on the Ettan IPGphor Important Before proceeding make sure the Ettan IPGphor unit is placed on a level surface Important Ensure that the three foam pads have been removed from the lid of the Ettan IPGphor um 80 6499 52 p6 Instructions O Rehydrate the IPG strips with the gel side down in the appropriate volume of rehydration solution using the Immobiline DryStrip Reswelling Tray Rehydration in the Cup Loading Manif
12. fold system Only one of each item shown A Manifold ceramic tray G Pre cut paper bridges B Pre cut electrode wicks H Sample cup insertion tool C Lid adapter I Cleaning solution D Electrode assembly J Forceps E Sample cups K Spirit level Fi Cleaning brush e Ettan IPGphor Cup Loading Manifold Unpacking Unwrap all packages carefully and compare contents with the packing list making sure all items arrived If any part s is missing contact your local Amersham Biosciences sales office Inspect all components for damage that may have occurred while the unit was in transit If any part s appear damaged contact the carrier immediately Be sure to keep all packing material for damage claims or to use should it become neces sary to return the unit Ettan IPGphor Cup Loading Manifold The Ettan IPGphor Cup Loading Manifold is suitable for first dimension isoelectric focusing IEF of proteins on Immobiline DryStrip immobi lized pH gradient IPG gel strips 3 mm wide from 7 cm to 24 cm long in the Amersham Biosciences Ettan IPGphor and Ettan IPGphor II Isoelectric Focusing Units IEF is performed with the gel side facing up Samples are applied in a localized region through an open bottom loading cup Cup loading has been found to improve protein focusing patterns particularly on basic IPG strips pH 6 9 and 6 11 Under conditions where substantial water transport electroendosmosis accompanies focusing such as
13. in the manifold through the paper wicks and or paper bridge parts Check that the IPG strip is fully rehydrated along its entire length Electrical contact at the electrodes is reduced by incomplete rehydration Check that the paper wicks are present and properly positioned Voltage too low or does not reach maximum set value Sample leaks from cup Ettan IPGphor protocol settings are incorrect for the experiment Conductivity ionic strength is too high Incorrect cup placement Incorrect strip placement Check that the current limit is properly set Check that the actual number of Immobiline DryStrip gels on the Ettan IPGphor platform is the same as the number of gels entered in the protocol Prepare the sample to yield a salt concentration less than 10 mM The recommended IPG Buffer concentration is 0 5 A maximum of 2 is advisable only if sample solu bility is a problem High conductivity can also arise from the use of poor quality urea or other denaturants Urea is also prone to decompose to charged breakdown products Higher conductivity salts and ionic impurities in the sample can raise the conductivity of the strip Shorter length IPG strips e g 7 cm strips will not reach 8000 V The distance between the electrodes is shorter so that the voltage gradient V cm required to reach the 50 pA current limit is reached at a lower overall voltage Check that the feet of the cups are resting on the bottom of the manifol
14. le cups A maximum of 150 ul of sample may be placed in these cups Check to make sure that there is cover fluid over the samples When the cups are initially placed on the manifold cover fluid will flow into the cups as they are seated When sample is introduced into the cups the sample will sink to the bottom of the cup and contact the IPG strip Note For basic IPG strips superior focusing patterns are generally obtained when the sample cup is placed as close to the anodic electrode as possible Close the Ettan IPGphor lid Program the Ettan IPGphor with the desired run parameters Ramping the voltage slowly while the sample is entering the IPG strip will improve results Optimal ramp voltages and times or Vhrs volt hours totals must be determined empirically for each sample type Focusing after sample cup application frequently requires fewer Vhrs than in gel sample rehydration loading methods particularly on basic pH range strips Important The Ettan IPGphor unit is capable of producing thousands of volts Before operating the unit read and fully understand the Ettan IPGphor operating instructions and warnings Ettan IPGphor Cup Loading Manifold suggested run conditions Suggested run conditions Table 1 Guidelines for Ettan IPGphor protocols with Cup Loading Manifold for broad medium and narrow pH range Immobiline DryStrip gels Running conditions Temperature 20 C Current 50 pA strip
15. n Dreieck soll den Benutzer auf die Anwesenheit wichtiger Betriebs und Wartungsanweisungen in der dem Ger t beiliegenden Dokumentation hinweisen Wenn Sie Anmerkungen zu diesem Handbuch haben dann senden Sie diese bitte an Amersham Biosciences AB SE 751 84 Uppsala Sweden Amersham Biosciences beh lt sich das Recht vor die Spezifikationen ohne vorhergehende Ank ndigung zu ndern Gew hrleistung and haftung Amersham Biosciences garantiert da das gelieferte Produkt sorgf ltig auf die Einhaltung der ver ffentlichten Spezifikationen getestet wurde Die in den Lieferbedingungen n her erl uterten Gew hrleistungsanspr che gelten nur dann wenn das Produkt gem den von Amersham Biosciences gelieferten Anweisungen installiert und benutzt wurde Amersham Biosciences bernimmt keinerlei Haftung f r Sch den oder Folgesch den einschlie lich aber nicht begrenzt auf Gewinneinbu en Einkommensverluste entgangene Gesch ftsab schl sse Verlust der Gebrauchsf higkeit oder andere Verluste die wie auch immer durch eine fehlerhafte oder unsachgem e Verwendung des Produkts verursacht wurden Die vorliegende Ver ffentlichung darf nur mit vorhergehender schriftlicher Genehmigung durch das Unternehmen vervielf ltigt in einem Abrufsystem gespeichert oder in irgendeiner Form oder mit irgendwelchen Mitteln bertragen werden piii Ettan IPGphor Cup Loading Manifold e safety warnings and precautions Italiano
16. ns aucun pr avis Garantie et responsabilit Amersham Biosciences garantit l utilisateur que le produit livr a subi avec succ s tous les essais pr vus pour s assurer qu il est conforme aux sp cifications et normes en vigueur La garantie incluse dans les conditions de livraison n est valable que si le produit a t install et utilis conform ment aux instructions fournies par Amersham Biosciences La soci t Amersham Biosciences ne sera en aucun cas responsable de tout dommage caus directe ment ou ndirectement par toute utilisation incorrecte ou non approuv e du produit ou d coulant de cette utilisation y compris toute perte de b n fice ou de recettes toute perte de perspectives commerciales tout emp chement d utilisation et tout autre risques ayant un rapport avec l utilisation du produit mais sans aucune limitation quant la nature de ces dommages La reproduction le stockage dans un syst me de r cup ration d informations ou la transmis sion sous quelque forme que ce soit et par quelque moyen que ce soit de la pr sente publi cation en totalit ou en partie sont strictement interdits sans autorisation pr alable crite de la soci t Espanol Deutsch e safety warnings and precautions Informacion importante para el usuario Para comprender el producto y utilizarlo con seguridad es necesario leer este manual en su totalidad El signo de admiraci n en un tri ngulo equil tero
17. old is not recommended the channel is too wide to ensure proper rehydration Follow the instructions included with the Immobiline DryStrips The table below is provided as a reference IPG strip Rehydration length cm volume ul 7 125 11 200 13 250 18 340 24 450 2 Cover the IPG strips with Immobiline DryStrip Cover Fluid and allow the strips to rehydrate overnight 10 20 hours Clean and dry the IPGphor bed before placing the manifold tray on the unit Position the manifold on the IPGphor platform The small T shaped protrusion fits into a cut out section of the IPGphor bed near the lid hinge Fig 10 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 unit Fig 11 Placement of IPG strips in manifold channels Note f cathodic cup loading is going to be used the strips should be placed such that the anodic end of the strips is 3 4 cm beyond the etched placement mark e instructions Q 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 Transfer the strips to the Ettan IPGphor Cup Loading Manifold Place the strips under the cover fluid face up in the tray with the anodic pointed end of the IPG strip resting on the appropriate mark etched into the bottom of the manifold ch
18. otal 7 05 24 0 9 30 28 0 Narrow 1 Step and Hold 300 3 00 0 9 3 00 0 9 intervals 2 Gradien 1000 6 00 3 9 6 00 3 9 3 Gradien 8000 3 00 13 5 3 00 13 5 4 Step and Hold 8000 5 10 41 7 6 00 47 7 Total 17 20 60 0 18 00 66 0 24 3 10 1 Step and Hold 300 3 00 0 9 3 00 0 9 3 10 NL 2 Gradien 1000 6 00 3 9 6 00 3 9 4 7 3 Gradien 8000 3 00 135 3 00 13 5 3 7 4 Step and Hold 8000 4 40 36 7 5 50 46 3 Total 16 40 55 0 17 50 65 0 6 9 1 Step and Hold 300 3 00 0 9 3 00 0 9 2 Gradien 1000 6 00 3 9 6 00 3 9 3 Gradien 8000 3 00 13 5 3 00 13 5 4 Step and Hold 8000 5 10 41 7 6 40 53 7 Total 17 10 60 0 18 40 72 0 Narrow 1 Step and Hold 300 3 00 0 9 3 00 0 9 intervals 2 Gradien 1000 6 00 39 6 00 3 9 3 Gradien 8000 3 00 13 5 3 00 13 5 4 Step and Hold 8000 9 40 77 7 11 40 93 7 Total 21 40 96 0 23 40 112 0 Narrow intervals 3 5 4 5 4 0 5 0 4 5 5 5 5 0 6 0 and 5 5 6 7 um 80 6499 52 p10 Troubleshooting first dimension IEF e troubleshooting Ettan IPGphor Isoelectric Focusing System employing the Cup Loading Manifold symptom possible cause remedy Current is too low or zero Electrical continuity is impeded Check the external electrode contacts The electrodes at the bottom of the strip holder one at each end must make metal to metal contact with the appropriate electrode contact area Check the internal electrode contacts The gel which becomes visible because of the dye in the rehydration solution must contact both electrodes
19. um 80 6499 52 piv Informazioni importanti per l operatore Per un utilizzo sicuro del prodotto leggere attentamente l intero contenuto del presente manuale Il punto esclamativo all interno di un triangolo equilatero indica all operatore la presenza di importanti istruzioni di funzionamento e manutenzione nella documentazione allegata al prodotto Si prega di inviare eventuali commenti al presente manuale a Amersham Biosciences AB SE 751 84 Uppsala Sweden Amersham Biosciences si riserva il diritto di apportare modifiche ai dati tecnici senza preavviso Garanzia e responsabilit Amersham Biosciences garantisce che prima della consegna il prodotto stato collaudato a fondo per soddisfare i requisiti specificati La garanzia inclusa nelle condizioni di consegna risulta valida solamente se il prodotto stato installato ed utilizzato nel rispetto delle istruzioni fornite da Amersham Biosciences Amersham Biosciences non potr essere ritenuta responsabile di incidenti o danni consequenziali inclusi ma non limitati a perdite di profitti mancato guadagno perdite di affari difetti di funzionamento e relative esposizioni dovuti ad un utilizzo non corretto del prodotto Nessuna parte della presente pubblicazione pu essere riprodotta conservata in sistemi di gestione dati o trasmessa in alcun forma n per nessuno scopo senza autorizzazione scritta del produttore Fig 1 Components of the Ettan IPGphor Cup Loading Mani
20. utes incorrect cup position N correct cup position Wi RTE Rio Rd TUTTO RUL oL IG N Fig 13 Correct placement of paper wicks Fig 14 Placement of electrode on paper wicks Cams are in the open position Fig 15 Placement of paper wick overlapping end of gel electrode on paper wick and sample cup on gel e instructions O Count out the appropriate number of precut paper wicks Two wicks per strip are required Separate the wicks from each other Add 150 pl distilled water to each wick Place the wicks on the IPG strips such that one end of the wick overlaps the end of the gel on the IPG strip Fig 13 The electrode must contact the wick With the electrode cams in the open position place the electrode assembly on top of all the wicks 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 Fig 14 O Briefly centrifuge the protein sample prior to loading to remove insoluble material and particulate matter These materials will impede sample entry and result in vertical streaks in the second dimension gel Load samples into the samp
21. with protein loads in excess of 1 mg the face up mode frequently yields better resolution The manifold can accommodate anodic and cathodic loading Ettan IPGphor Cup Loading Manifold Ettan IPGphor Cup Loading Manifold forcep wells channel numbering centering protrusion Fig 2 Ceramic manifold with details cam in open position cam in closed position Fig 3 Electrode and details um 80 6499 52 p2 Manifold The manifold Fig 2 is made of aluminum oxide ceramic for efficient heat transfer and temperature control during IEF Protrusions along the channel inside the manifold align the rehydrated IPG strip keeping it straight and centered when placed inside the manifold The manifold has a coating to minimize protein adsorption Because some cleaning agents can remove this coating clean the manifold only with the Ettan IPGphor Strip Holder Cleaner The manifold is fragile and should be handled with care IPG strips from 7 cm to 24 cm long may be used The unit may also be used for IPG strip equilibration prior to second dimension electro phoresis Forcep wells are located at the ends of the channels to aid strip removal after electrophoresis Electrode Assembly The movable electrodes Fig 3 can be placed anywhere along the mani fold where the electrode pins will make electrical contact with the power supply pads on the bed of the Ettan IPGphor The electrodes have a plat inum wire that is strung
22. yStrip Reswelling Tray 7 24 cm 1 80 6465 32 Immobiline DryStrip Cover Fluid 1 liter 17 1335 01 Ettan IPGphor Strip Holder Cleaner 950 ml 80 6452 78 p13 Ettan Immobiline IPGphor and PlusOne are trade marks of Amersham Biosciences Limited Amersham and Amersham Biosciences are trademarks of Amersham plc Amersham Biosciences Corp 2003 All rights reserved All goods and services are sold subject to the terms and conditions of sale of the company within the Amersham Biosciences group that supplies them A copy of these terms and conditions is available on request Printed in the USA www amershambiosciences com Amersham Biosciences UK Limited Amersham Place Little Chalfont Buckinghamshire England HP7 9NA Amersham Biosciences AB SE 751 84 Uppsala Sweden Amersham Biosciences Corp 800 Centennial Avenue PO Box 1327 Piscataway NJ 08855 USA Amersham Biosciences Europe GmbH Munzinger Strasse 9 D 79111 Freiburg Germany Amersham Biosciences
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