TE70X, TE70XP, TE77X and TE77XP User Manual
Contents
1. If any part appears damaged contact the carrier immediately Be sure to keep all packing material for damage claims or to use should it become necessary to return the unit TE70X TE77X TE70XP TE77XP This declaration of conformity is only valid for the instrument when it is e used in laboratory locations e used as delivered from Hoefer Inc except for alterations described in the user manual e connected to other CE labeled instruments or products recommended or approved by Hoefer Inc Specifications Electrode size cm 14x 16 Inputs Voltage V 30 DC Current A 0 5 Output Voltage VDC 30 Current A 0 5 Frequency Hz 21 x 26 14x 16 21 x 26 30 DC 100 240 AC 100 240 AC 0 5 0 7 0 7 30 30 30 0 5 0 5 0 5 47 63 47 63 Maximum setting for the customer s power supply Minimum transfer stack thickness TE70X TE70XP TE77X TE77XP Environmental operating conditions Dimensions w x d x h Weight Certification 3 2 mm 12 0 mm 2 8 mm 12 0 mm Indoor use 4 40 C Humidity up to 80 Altitude up to 2000 m Installation category II Pollution degree 2 38 x 46 x 9 cm 15 x 18 x 3 5 in Shipping 6 8 kg Unit 3 7 kg EN61010 1 EN 61326 CE UL61010 1 2004 CSA 22 2 61010 1 04 Note Take care to place the gel correctly on the first try because proteins begin to transfer immediately once transfer has begun moving the gel will distort results or2. 10 7 1 10 7 3 Greene Publish ing and Wiley Interscience NY 1991 Smejkal G and Gallagher S Determination of Semidry Protein Transfer Efficiency with Transverse Gradient Gel Electrophoresis Biotechniques 16 196 202 1994 Tovey E and Baldo B Comparison of semi dry and conventional tank buffer electrotransfer of proteins from polyacrylamide gels to nitrocellulose membranes Electrophoresis 8 384 387 1987 e pl9 Important Request a copy of the Hoefer Inc Health and Safety Declaration form before returning the item No items can be accepted for servicing or return unless this form is properly completed Note A Return Authorization RA number must be obtained from Hoefer Inc before returning any item to Hoefer Inc p20 7 Customer service information Technical service and repair Hoefer Inc offers complete technical support for all of our products If you have any ques tions about how to use this product or would like to arrange to repair it please call or fax your local Hoefer Inc representative Check the Hoefer Inc website at www hoeferinc com for the distributor in your area Or contact us directly at Hoefer Inc 84 October Hill Road Holliston MA 01746 Toll Free 1 800 227 4750 Phone 1 508 893 8999 Fax 1 508 893 0176 support hoeferinc com www hoeferinc com 8 Ordering information product qty code no TE70X Semi
3. and buffer can be added to the transfer stack Replace cover and press any key except Power ON OFF to continue For future transfers add more layers of buffer soaked blot paper e pll Note Staining the gel s for residual protein gives an indication of the completeness of transfer Note Rewet dried membranes in the appropriate wetting buffer prior to processing e pl2 After the transfer is complete TE70X and TE77X Turn off the power supply and disconnect the leads from the power supply TE70XP and TE77XP After a transfer is complete the unit will beep for 5 seconds The LED display will blink and cycle between the values for the current time and voltage at the time the transfer was stopped These can be recorded into a notebook if desired Pushing any button will clear the display 0 Remove the cover slowly because the stack may adhere to it e Remove and dispose of the upper blot papers Remove the gel s Optional Stain gel to check for residual protein left in the gel o If desired label the gel contact side of the membrane with a soft pencil Remove the membrane s from the stack with blunt forceps Process the membrane according to your protocol or allow the membrane to air dry prior to storage Remove the remaining blot papers and dispose o Rinse the unit according to the Care and maintenance instructions next page 3 Care an
4. blot papers in future transfers No output voltage Note For more troubleshooting ideas refer to Bjerrum O J et al 1988 Transfer stack is being short circuited e p16 Note Buffers containing methanol may deteriorate if stored for long periods add methanol just prior to transfer 5 Electrotransfer notes e Run the transfer as soon as possible after electrophoresis to minimize protein diffusion within the gel Stacked gels must all be the same size Limit transfers to two hours or less The recommended methanol concentration for different membrane types are membrane type methanol Charged nylon 0 Nitrocellulose 10 20 PVDF 10 20 e Use a buffer with low ionic strength such as one of the two listed below to prevent over heating Use the CAPS buffer when Tris cannot be used e g peptide sequencing CAPS can improve transfer because of its effect on the charge of the protein see Matsudaira 1987 Towbin buffer 25 mM Tris 192 mM glycine 20 wv methanol pH 8 3 1 liter Tris FW 121 1 25 mM 3 0g Glycine FW 75 07 192 mM 14 4 g SDS FW 288 4 0 1 3 5 mM 1 0g Dissolve in 600 ml distilled water Add methanol as required Bring to 1 liter with distilled water Do not adjust the pH which should be between 8 2 8 4 Optional Chill before use Optional Adding SDS can improve transfer efficiency iDepending on the membrane type selected see table above adding metha
5. cause shadow bands on the blot 2 Operating instructions To transfer proteins prepare the unit assemble the stack and connect to a power supply if necessary Then run the transfer for the required amount of time Each step is described below 0 Prepare the unit by rinsing the electrodes with distilled water a Prepare the gel Cut away the wells and or stacking gel section Eguilibrate the gel in transfer buffer if reguired Prepare the transfer stack Cut the blot paper and transfer membrane to the same size as the gel Stack the layers carefully so the edges align If for some reason the membrane needs to be larger than the gel use a mylar mask see optional step below to ensure the current does not bypass the gel Optional Cut a mask to the proper size Measure the gel Cut an opening centered in the solid mask to a size roughly 2 mm smaller than the gel Take care with sharp blades when cutting the masks Place the mask in the base of the unit centering the opening Note Each 21 x 26 cm blot paper will absorb approximately 50 ml of transfer buffer Each 14 x 16 cm blot paper will absorb approximately 20 ml of buffer Note If not using Hoefer blotting paper additional pieces may be required to achieve desired results Note Always wear gloves when handling membranes to avoid leaving fingerprints o Prepare the blotting paper For each gel cut at least 6 pieces of blo
6. d rfen sondern separat behandelt werden miissen Bitte nehmen Sie Kontakt mit einem autorisierten Beauftragten des Herstellers auf um Informationen hinsichtlich der Entsorgung Ihres Ger tes zu erhalten Questo simbolo indica che i rifiuti derivanti da apparecchiature elettriche ed elettroniche non devono essere smaltiti come rifiuti municipali indifferenziati e devono invece essere raccolti separatamente Per informazioni relative alle modalita di smantellamento delle apparecchiature fuori uso contattare un rappresentante autorizzato del fabbricante Este simbolo indica que el equipo el ctrico y electr nico no debe tirarse con los desechos dom sticos y debe tratarse por separado Contacte con el representante local del fabricante para obtener mas informaci n sobre la forma de desechar el equipo Denna symbol anger att elektriska och elektroniska utrustningar inte far avyttras som osorterat hushallsavfall och maste samlas in separat Var god kontakta en auktoriserad tillverkarrepresentant f r information angaende avyttring av utrustningen e pvii TE70X top electrode voltage leads lower electrode RESET circuit breaker reset TE77XP power top electrode cords power lower electrode transformer Fig 1 Semi dry transfer unit main components 1 TE70X Semi dry Blotters Description The TE70X TE77X TE70XP and the TE77XP semi dry blotters rapidly transfer proteins from polyacrylamide gels ont
7. dry Transfer Unit 14 x 16 cm 1 TE7OX Includes 25 sheets of blotting paper 50 sheets of cellophane and 2 solid masks TE77X Semi dry Transfer Unit 21 x 26 cm 1 TE77X Includes 25 sheets of blotting paper 50 sheets of cellophane and 2 solid masks TE7OXP Semi dry Transfer Unit 14 x 16 cm 1 TE70XP Includes 25 sheets of blotting paper 50 sheets of cellophane and 2 solid masks TE77XP Semi dry Transfer Unit 21 x 26 cm 1 TE77XP Includes 25 sheets of blotting paper 50 sheets of cellophane and 2 solid masks Accessories TE70X and TE70XP Solid masks 16 5 x 18 5 cm 4 TE74 Porous cellophane 20 x 35 5 cm 50 TE73 Blotting paper precut 14 x 16 cm 25 TE76 1416 TE77X and TE77XP Solid masks 23 x 27 5 cm 4 TE78 Porous cellophane 35 x 44 cm 50 SE1142 Blotting paper precut 21 x 26 cm 25 TE76 Power Cord Kit 1 set PS36 24 Fuse Micro 1 6 A 250 V 5 PSF1 6A MICRO e p21 Related products product code no SE600 Chroma SE600X 15 1 5 MiniVE Vertical Electrophoresis system SE300 10A 1 0 PBS300B Power Supply 300 V 500 mA 90 W PBS300B Transfer Membranes Pure Nitrocellulose sheets and roll 0 45 um pore size 8 x 9 5 cm 10 sheets GM NC45 89 16 x 16 cm 10 sheets GM NC45 1616 20 x 23 5 cm 10 sheets GM NC45 2320 30 cm x 3 m 1 roll GM NC45 0 2 um pore size 30 cm x 3 m 1 roll GM NC22 PVDF membrane 0 45 um pore size 10 x 10 cm 10 sheets GM PV45 1010 15 x 15 cm 5 sheet
8. elektrick bezpe nostn normy P esto e by m ly b t pou ity pouze dn vy kolen oper tor st cel toto ru n p ed pou it m n stroje a pou it pouze v souladu s pokyny P stroj mus b t v dy pou vaj se na v konu zemi v st ra spr vn zemn ny k zemi na s ti v stce Vyu it pouze nepo kozen elektrick dr ty a vybaven pro nap t budete pou vat V echna za zen spojen s vysok m nap t m by m la b t v souladu s EN61010 1 2001 Si ponech n stroje jako such a ist jako mo n Ot ete pravideln s a m kk vlhk m had kem Nech je n stroj nenastaven pln p ed pou it m Nejsou provozov na na n stroj v extr mn vlhkost nad 80 P ede lo kondenzaci o pron jmu jednotky na okoln teplotu nech p i p ij m n n stroj z chladn j do tepl ho prost ed Pro umo n n dostate n chlazen zajistit aby otvory n stroje jsou nevztahuje Vigtig Information Danish Hvis dette udstyr bruges i en made ikke specificeret ved Hoefer Inc den beskyttelse som er blevet forsynet af udstyret kan m ske svaekkes Dette instrument er designet for indendors laboratori umbrug bare Bare tilbeh r og del godkendede eller forsynede ved Hoefer Inc kan m ske bruges for drive funktionsfejl og betjening dette produkt Advare Fordi dette instrument kan udvikle tilstraek kelig spaending og strom at fremstil
9. i cz ci zatwierdzone lub dostarczone przez Hoefer Inc mog by wykorzystane do eksp loatacji utrzymania i obs ugi tego produktu Uwaga Poniewa ten akt prawny mo e by rozwini cie odpowiednich napi cie i bie cych do wyprodu kowania miertelnego szoku opiek musi by wykony wane w dzia aniu Ten instrument zosta zaprojektowany zgodnie z tym EN61010 1 2001 Bezpiecze stwo elektryczne standard Niemniej jednak nale y stosowa jedynie przez odpowiednio przeszkoleni operator w Znajd pa stwo to ca y podr cznika przed zastosowaniem instrumentu i stosowa jedynie zgodnie z instrukcjami Instrument musi zawsze by wykorzystane z ziemi doprowadzi do zasilania detonuj cego w a ciwie uzasadnione na ziemi w sieci wodoci gowej rynku zbytu Wykorzystanie tylko nieuszkodzona elektrycznych drut w i urz dzenia specjalne do napi zap ac wykorzystania Wszystkie urz dzenia pod czone do wysokiego napi cia powinny by zgodne z EN61010 1 2001 Kontrolowa instrumentu jako suche i czyste jak to mo liwe Wytrze regularnie przy pomocy mi kkiego wilgotnej szmatki Niech si instrumentem ca kowicie wysuszy przed u yciem Nie prowadz do instrumentu w skrajnych wilgotno ci powy ej 80 Zapobiec kondensacji najmu przez jednostk r wnowa y do temperatury pokojowej przy podejmowaniu instrumentu z ch odniejsze w cieplejszych rodowiska Aby umo liwi wystarczaj ce c
10. or 1 2 mm smaller Larger sizes will provide an electrical path for current to bypass the gel solution Different proteins will transfer at different rates depending on size and net charge e p15 problem solution Inefficient binding to membrane Chemical parameters Prepare transfer buffer without SDS SDS can improve transfer efficiency but can interfere with protein binding to a PVDF membrane Add 10 20 methanol to the transfer buffer to enhance binding to nitrocellulose or PVDF Membrane parameters Use a membrane with a smaller pore size 0 20 um if proteins pass through the membrane Place a membrane both over and under the gel to capture any proteins migrating in the opposite direction Check if too much sample is available for the binding surface area by placing two membranes instead of one If blow through occurs reduce the sample load Wear gloves when handling membranes Store membranes at ambient temperature and out of direct sunlight Check shelf life of nitrocellulose membrane replace if necessary TE70XP TE77XP Current reading lower than setting Maximum voltage 30 V has been reached Buffer may be depleted No output current Transfer stack is not making contact with the upper electrode Add buffer soaked blot papers dry error message Instrument has detected large changes in transfer stack resistance Add more buffer soaked
11. paper additional pieces may be c Cover the gel with three layers of buffer saturated reguired to achieve desired results blotting paper Fig 2 Transfer stack for a single gel Blotting paper 2 3 sheets These stack Gel components are the same size Membrane as the gel or slightly smaller Blotting paper 2 3 sheets Mask opening approx 2 mm smaller than the gel on all sides Optional p Note When ge on ge such fact position buffer mem im portantly ransferring multiple s transfer efficiency depends ors as gel thickness in the stack transfer brane type and most the characteristics of the protein The gel closest to the anode generally transfers the most completely ge t is preferable to lay s side by side rather than stacking them Fig 3 Transfer stacks for gels placed side by side Note There must be no electrical contact between the two stacks Multiple gels Either lay gels of the same thickness side by side Fig 3 or stack 2 sandwiches layered as shown Fig 4 For best results the transfer stack should be centered in the electrode panels If two gels are stacked separate them with porous cellophane not plastic wrap Cellophane permits electric current to pass but stops proteins Cut the cellophane slightly smaller than the gel and wet with transfer buffer Several sheets of buffer soaked blotting paper on each gel prov
12. Ne pas fonctionner l instrument dans l extr me humidit au dessus de 80 Eviter la condensa tion en laissant l equilibrate d unit la temp rature ambiante en prenant l instrument d un plus froid un environnement plus chaud Permettre le refroidissement suffisant garantir que les conduits de l instrument ne sont pas couverts Wichtige Informationen German Wenn diese Ausr stung gewisserma en nicht angege ben durch Hoefer Inc verwendet wird kann der durch die Ausriistung zur Verfiigung gestellte Schutz verschlechtert werden Dieses Instrument wird f r den Innenlaborgebrauch nur daf r entworfen Nur Zus tze und Teile genehmigten oder lieferten durch Hoefer Inc kann fiir das Funktionieren das Aufrechterhalten und die Wartung dieses Produktes verwendet werden Die Warnung Weil dieses Instrument gen gend Strom spannung und Strom entwickeln kann um einen t dli chen Sto zu erzeugen muss Sorge in seiner Operation ausge bt werden Dieses Instrument wird in bereinstimmung mit dem EN61010 1 2001 elektrischen Sicherheitsstandard daf r entworfen Dennoch sollte es nur von richtig erzo genen Maschinenbedienern verwendet werden Lesen Sie dieses komplette Handbuch vor dem Verwenden des Instrumentes und verwenden Sie nur gemaB den Instruktionen Das Instrument muss immer mit der Erdleitung der Macht Schnur richtig niedergelegt zur Erde am Hauptausgang verwendet werden Nur unbesch dig
13. TE70X Semi dry Blotters TE7OX Semi dry transfer unit TE77X Semi dry transfer unit TE7OXP Semi dry transfer unit TE77XP Semi dry transfer unit um TE70X IM Rev CO 07 12 H O e fe r Contents Important INFORMATION lt ssassasisiassmaaskaaa kan ze de ji Waste Electrical and Electronic Equipment WEEE ee vii 1 TE7OX Semi dry Blotters Description 1 Unpacking aaoi se ee ege ee 2 Specifications 3 2 Operating instructions irsz ennnen 4 Electrotransief sadow esnean ek 9 3 Care and Maintenance ssa szwy ES dE 13 A TrOUDIESMOORING ae O A W PA AAAA tees 14 5 Electrotranstey notes siew Ea 17 6 Bib OSTA PAY repineen kananan er 19 7 Customer service information 1 111 20 8 Ordering informatio ranees 21 Important Information English If this equipment is used in a manner not specified by Hoefer Inc the protection provided by the equipment may be impaired This instrument is designed for indoor laboratory use only Only accessories and parts approved or supplied by Hoefer Inc may be used for operating maintaining and servicing this product Warning Because this instrument can develop suffi cient voltage and current to produce a lethal shock care must be exercised in its operation This instrument is designed in accordance with the EN61010 1 2001 electrical safety standard Never theless it should be used only by properly trained operators Read t
14. avend en onderhouden van dit product Waarschuwend Omdat dit instrument voldoende spanning en stroom kan ontwikkelen om een dodeli jke schok te produceren moet zorg in zijn operatie worden geoefend Dit instrument is in overeenstemming met de EN61010 1 2001 elektrische veiligheidsstandaard ontworpen Niettemin zou het enkel door goed getrainde bedieningslieden moeten worden gebruikt Lees dit volledige handboek voor het gebruik het instrument en gebruik enkel volgens de instructies Het instrument moet altijd met de aardeleiding van het stroomsnoer correct grondde naar aarde aan het hoofdafzetgebied worden gebruikt Gebruik enkel onbeschadigde elektrische draad en uitrustings specifiek voor de spanningen u zult gebruiken Alle uitrustingen sloten aan aan hoogspan ning zou in overeenstemming met EN61010 1 2001 moeten zijn Houd het instrument zo droge en schone zoals mogelijk Bij Wis regelmatig met een zacht temper doek Verhuur het instrument droogt volledig voor het gebruik Bedien niet het instrument in extreme vochtigheid bovenstaande 80 Vermijd condensatie door het verhuren van de eenheid in evenwicht brengt naar omgevingstemperatuur wanneer nemen het instru ment van een kouder naar een lievere omgeving Om toe te staan voldoende afkoelen verzeker dat de luchtopeningen van het instrument niet bedekt zijn T rke Tietoa Finnish Jos t t varusteita k ytet n tavassa ei m ritetty H
15. d maintenance e Do not autoclave or wash the unit in a dishwasher e Do not immerse the unit in water Rinse the cathode in the cover and the anode in the base with distilled water Let the unit air dry completely If using radioactive reagents decontaminate the unit with a cleaning agent such as Contrad 70 Never use abrasive cleansers e pl3 4 Troubleshooting problem solution Incomplete transfer Blank or faint areas on Remove trapped air pockets between the gel and the membrane membrane during stack assembly Use buffer with a lower ionic strength Molecules do not migrate Check all electrical connections Confirm that current out of gel is flowing through the transfer stack Check that the buffer pH is close to the intended pH Most buffers should not be titrated Make fresh buffer Use 3 5 mM SDS 0 1 in the transfer buffer Add several more sheets of buffer saturated blotting paper to each side of the gel sandwich so that more buffer is present during the transfer Increase the transfer period Large fragments may require an additional hour Do not use staining or fixing agents on the gel before transfer Use a thinner gel Reduce the gel acrylamide concentration If using a non nitrocellulose membrane avoid includ ing methanol in the transfer buffer or reduce the amount to the minimum possible Use reagent grade chemicals Increase the net charge on the prote
16. du ska anv nda All utrustning kopplats som till h g spanning skulle vara i verensst mmelse med EN61010 1 2001 Haller instrumentet d torkar och reng r som m jlig Torka regelbundet med en mjuk fuktig trasa Lat instrumentet torka fullstandigt fore anvandningen Fungerar inte instrumentet i extrem fuktighet ver 80 Undvik kondensering vid lata enheten equili brate till omgivande temperatur nar ta instrumentet fran en kallare till en varmare milj Att till ta tillr cklig kyla ser till att halen av instru mentet inte tacks o pvi English Eas French R muu German R Ea Italian R E Spanish La Swedish R Waste Electrical and Electronic Eguipment WEEE This symbol indicates that the waste of electrical and electronic eguipment must not be disposed as unsorted municipal waste and must be collected separately Please contact an authorized representative of the manufacturer for information concerning the decommissioning of your eguipment Ce symbole indigue que les d chets relatifs a guipement lectrique et lectronique ne doivent pas tre jet s comme les ordures m nag res non tri es et doivent tre collect s s par ment Contactez un repr sentant agr du fabricant pour obtenir des informations sur la mise au rebut de votre quipement Dieses Symbol kennzeichnet elektrische und elektronische Ger te die nicht mit dem gew hnlichen unsortierten Hausm ll entsorgt werden
17. et Varsler Fordi dette instrumentet kan utvikle tilstrek kelig spenning og strom til produsere et dodelig sjokk m bli vd bekymring i dets drift Dette instrumentet er utformet i samsvar med EN61010 1 2001 elektrisk sikkerhetsstandard Likevel burde bli brukt det bare av skikkelig utdannede oper atorer Les denne hele handboken for bruking instru mentet og bruken bare gi til instruksjonene Instrumentet m alltid bli brukt med jorden blyet av kraftkabelen som riktig ha blitt jordet til jord pa hovedledningen utlop Bruker bare uskadd elektrisk ledningsfremforing og utstyr som er spesifikk for spenningene du vil bruke All utstyr koplet til hoyspenning burde veere i samsvar med EN61010 1 2001 Beholder instrumentet som t rker og rengjor som mulig Visk regulaert med et mykt fuktig stoff La instru mentet torker komplett for bruk Driver instrumentet i ekstrem fuktighet ikke ovenfor 80 Unnga kondensasjon ved a la enheten equili brate til omgivelsestemperatur ved taen instrumentets fra et kaldere til et varmere miljo Til tillate tilstrekkelig kj lig sikrer at ventilasjon sapningene av instrumentet er ikke dekket Wazne Informacje Polish Je eli ten sprz t jest wykorzystywany w spos b nie okre lone przez Hoefer Inc do ochrony przewidzianej przez urz dzenie mo e zosta obni ony Instrument ten jest przeznaczony do u ytku w labora toriach kryty tylko Tylko akcesori w
18. h odzenia zapewniaj e rozci cia of the instrument nie obj te ubezpieczeniem Informac es Importantes Portuguese Se este equipamento usado numa maneira nao espe cificada por Hoefer Inc que a protecc o fornecida pelo equipamento pode ser comprometida Este instrumento projectado para uso de interior de laborat rio s S acess rios e partes aprovaram ou forneceu por Hoefer Inc pode ser usada para operar manter e servicing este produto Advertindo Porque este instrumento pode desen volver voltagem suficiente e corrente produzir um choque letal cuidado deve ser exercitado em sua operac o o pv Este instrumento projectado de acordo com o EN61010 1 2001 condic o de seguranca el ctrica Nao obstante deve ser usado s por operadores adequada mente treinados Leia este manual inteiro antes de usar o instrumento e use s de acordo com as instrucc es O instrumento sempre deve ser usado com o chumbo de terra do cord o de poder corretamente baseou a terra nos canos sa da principais Usa fio el ctrico s intacto e equipamento espec fico para as voltagens que voc usar Todo equipamento conectado a voltagem alta deve ser de acordo com EN61010 1 2001 Mantem o instrumento tao seco e limpo como poss vel Limpe regularmente com um pano h mido macio Deixe o instrumento secar completamente antes de uso Nao opera o instrumento em humidade extrema acima de 80 Evite c
19. his entire manual before using the instrument and use only according to the instructions The instrument must always be used with the earth lead of the power cord correctly grounded to earth at the mains outlet Use only undamaged electrical wire and equipment specific for the voltages you will use All equipment connected to high voltage should be in accordance with EN61010 1 2001 Keep the instrument as dry and clean as possible Wipe regularly with a soft damp cloth Let the instru ment dry completely before use Do not operate the instrument in extreme humidity above 80 Avoid condensation by letting the unit equilibrate to ambient temperature when taking the instrument from a colder to a warmer environment To permit sufficient cooling ensure that the vents of the instrument are not covered Dulezit Informace Czech Pokud by toto za zen je pou ito zp sobem kter neni podle Hoefer ochrana poskytovan na z klad Inc za zen m e b t naru ena Tento n stroj je ur en pro vnit n pou it v laborato i pouze Pouze p slu enstv a sti schv len nebo poskyt nut ch Hoefer Inc mohou b t pou ity pro provoz dr bu a dr b tohoto v robku Pozor Proto e tento n stroj m e vyvinout dostate n nap t a proud kter m vyr b t a smrtiacej ok p e mus b t vykon v na v jeho provoz Tento n stroj je ur en v souladu s EN61010 1 2001
20. i sicurezza elettrica Tuttavia dovrebbe essere usato degli opera tori solo correttamente addestrati Leggere questo manuale intero prima di usare lo strumento e l uso solo secondo le istruzioni Lo strumento deve essere sempre usato col piombo di terra della spina di alimentazione correttamente hanno messo a terra alla terra alla presa di corrente principale Usa il filo metallico e apparecchiatura solo intatti elettrici specifici per i voltaggi che lei usera Tutta Vapparecchiatura collegata all alto voltaggio dovrebbe essere conformemente a EN61010 1 2001 Tiene lo strumento come secco e pulito come possibile Pulire regolarmente con un morbido per spegnere il panno Lasciare lo strumento asciuga completamente prima dell uso Non opera lo strumento nell umidita estrema al di o piv sopra di 80 Evitare la condensazione lasciando l unit equilibra alla temperatura ambiente quando portare lo strumento da un piu freddo a un ambiente piu caldo Di permettere raffreddare sufficiente assicura che gli sbocchi dello strumento non sono coperti Viktig Informasjon Norwegian Hvis dette utstyret blir brukt i en mate ikke spesifisert ved Hoefer Inc beskyttelsen som ha blitt git av utsty ret kan bli svekket Dette instrumentet er utformet for innendors labora toriumbruk bare Bare tilbeh r og deler godkjente eller forsynte ved Hoefer Inc kan bli brukt for drive vedlikeholde og betjene dette produkt
21. ides electrical continuity o Place the Cover on top of the transfer stack Do not remove the cover until after the transfer is complete in order to prevent stack components from moving Blotting paper 2 3 sheets Gel Membrane Blotting paper 2 3 sheets Mask optional Fig 4 Transfer stack for stacked gels Separate the stack for each gel with a sheet of cellophane Blotting paper 2 3 sheets Gel Membrane Blotting paper 1 sheet Cellophane Gel Membrane Blotting paper 2 3 sheets Mask optional Note Transfers exceeding one hour may require additional sheets of buffer saturated blot paper in the transfer stack Note Generally smaller fragments transfer more quickly than larger ones Fig 5 Recommended current settings for different gel sizes Use the graph to quickly find the current setting for your gel size or calculate the gel area cm and multiply it by 0 8 mA Electrotransfer TE70X and TE77X 0 The power supply should be switched off and both the current and voltage controls set at zero Then plug the color coded leads from the base of the transfer unit into the power supply jacks matching red to red and black to black Do not reverse polar ity The lid contains the black or negative electrode The base contains the red or positive electrode e Set the power supply voltage and current Set the voltage limit to 30 V maximum The i
22. ilmarei t peitet n Information Importante French Si cet quipement est utilis dans une mani re pas sp cifi par Hoefer Inc la protection fourni par V guipement pourrait tre diminu e Cet instrument est con u pour l usage de laboratoire int rieur seulement Seulement les accessoires et les parties ont approuv ou ont fourni par Hoefer Inc pourrait tre utilis pour fonctionner maintenir et entretenir ce produit Avertissant Parce que cet instrument peut d velopper la tension et le courant suffisants pour produire un choc mortel le soin doit tre exerc dans son op ration Cet instrument est conform ment con u a l EN61010 1 2001 norme de s curit lectrique N anmoins piii il devrait tre seulement utilis par les op rateurs convenablement entrain s Lire ce manuel entier avant utiliser l instrument et usage seulement selon les instructions Linstrument toujours doit tre utilis avec l avance de terre du cordon d alimentation correctement a fond a la terre a la sortie principale Utiliser le fil et l quipement lectriques seule ment intacts sp cifiques pour les tensions que vous utiliserez Tout guipement connect a haute tension devrait tre conform ment a EN61010 1 2001 Garder l instrument aussi sec et propre comme possi ble Essuyer r guli rement avec un doux touffer du tissu Laisser instrument s che completement avant l usage
23. in by using a transfer buffer with a different pH Lower pH lt 6 7 increases the positive charge on proteins higher pH gt 6 7 increases the negative charge on proteins e pl4 problem solution Open circuit or no output current for TE70X or TE77X Push circuit protection reset button located next to the input leads To prevent recurrence limit the input voltage to 30 V and the current to 500 mA Early models Blown fuse in protection circuit Replace 1 6 A micro fuse See Accessories in Ordering information Smeared or diffuse band patterns If equilibrating before the transfer shorten or elimi nate the equilibration time and or equilibrate under coldroom conditions If the transfer buffer contains gt 10 methanol equili brate the gel in transfer buffer for 30 minutes to allow it to shrink before assembling the stack Note Large proteins may not migrate as readily once the pore size is slightly reduced Take care that the gel does not shift once it contacts the membrane Buffer depletion can change the pH in the transfer stack and have a negative effect on the transfer On subsequent transfers either shorten the transfer time reduce the current or increase the number of buffer soaked blotting papers in the stack Check that the preferred binding surface of the membrane if any contacts the gel Uneven band transfer The blotting paper and membrane must be the same size as the gel
24. le et dadbringende chok skal pleje bruges i dets drift Dette instrument er designet i overensstemmelse med EN61010 1 2001 elektrisk sikkerhedstandard Alligevel skulle det bruges bare af passende traenede operatorer Laes denne hel handbog for bruging instrumentet og brug bare i henhold til instruktionerne Instrumentet skal altid bruges med jordblyet af netledningen rigtigt jordede til jord pa hovedledning sudlobet Bruger bare uskadt elektrisk trad og udstyr som v re specifik for spaendingerne du vil bruge Alt udstyr forbundet til haj spaending skulle vaere i overensstem melse med EN61010 1 2001 Beholder instrumentet s tor og ren som mulig Tor regul rt med et bl dt fugtigt stof Lad instrument torken komplet for brug Driver ikke instrumentet i yderst fugtighed ovenfor O pii 80 Undga kondensation ved lade enheden equili brate til omgivende temperatur ved tageen instru mentets fra et koldere til et varmere miljo At tillade tilstr kkelig afk ling forsikrer at lufthullerne af instrumentet er ikke d kket Belangrijke Informatie Dutch Indien deze uitrusting in een manier wordt gebruikt die niet door Hoefer is gespecificeerd Nv de bescherming die door de uitrusting is verzorgd kan worden geschaad Dit instrument is voor binnenlaboratoriumgebruik enkel ontworpen Enkel onderdelen en delen keurden goed of leverden door Hoefer Nv kan voor het bedienen worden gebruikt handh
25. nol can improve transfer results e p17 e p18 CAPS buffer 1X 10 mM CAPS pH 11 0 1 liter CAPS FW 221 3 10 mM 228 3 cyclohexylamino 1 propanesulfonic acid Dissolve in 600 ml distilled water adjust to pH 11 0 with conc NaOH Adjust volume to 1 0 liter e For a 3 buffer system refer to Kyhse Anderson J 1984 e Transfer efficiency varies depending on the gel concentration which can be optimized For more information refer to Smejkal and Gallagher 1994 6 Bibliography Bjerrum O J Larsen K and Heegaard N CRC Handbook of Immunoblotting of Proteins 1 Section 7 CRC Press 1988 Gallagher S Winston S E Fuller S A and Hurrell J G R Immunoblotting and Immunodetection In Current Protocols in Molecular Biology 10 8 1 10 8 17 Greene Publishing and Wiley Interscience NY 1993 Hancock K and Tsang V India ink staining of proteins on nitrocellulose paper Anal Biochem 133 157 162 1983 Kyhse Anderson J Electroblotting of multiple gels A simple apparatus without buffer tank for rapid transfer of proteins from polyacrylamide to nitro cellulose J Biochem and Biophys Meth 10 203 209 1984 Matsudaira P Sequence from Picomole Quantities of Proteins Electroblotted onto Polyvinylidene Difluoride Membranes J Biol Chem 262 10035 1987 Sasse J and Gallagher S Detection of Proteins on Blot Transfer Membranes In Current Protocols in Molecular Biology
26. nput voltage must not exceed 30 V or the protection will be activated The maximum current should not exceed 0 8 mA cm of the gel surface or 500 mA maximum If transferring several layers of gels a longer time may be required If available set the power supply timer Most transfers are complete within one hour but larger proteins proteins from native gels and thicker gels may require an additional 1 hour of transfer time The optimum transfer time for each protein and gel system must be determined empirically A cm2 500 Gel Size op J 300 200 o EI gt E o 8 H s o R s o a o Note While setting the time there is an extra setting labelled ON above the 4 00 upper limit This will disable the timer allowing the transfer to run continuously until manually turned off by the user Fig 6 TE7OXP keypad H transfer gt start stop up down arrows adjust value e plo TE70XP and TE77XP 0 Turn the instrument on using the POWER button on the front keypad The instrument works by setting the current and time and then starting the transfer During the transfer the voltage can be displayed The transfer cannot be set to run at constant voltage The DISPLAY MODE BUTTON toggles between the current mA time hour minutes and the voltage V The UP and DOWN arrows to change the value for the current and time e Press the START STOP button
27. o a membrane by means of a low current and low voltage electrotransfer with minimal Joule heating Most transfers are complete in one hour or less The smaller TE70X unit transfer surface is 14 x 16 cm suitable for transferring standard gels including those from the SE600 Chroma and the SE400 The larger TE77X transfer surface is 21 x 26 cm suitable for transferring large format gels The TE70X and TE77X have safety circuits built into the instrument that limit the voltage to 30 V and the current to 0 5 A The circuits protect the user from unnecessary electrical hazards The TE70XP and the TE77XP have a built in power supply for transferring gels This eliminates the need for an external power supply These instruments can deliver up to 30 V and 0 5 A The instruments also monitor the transfer stack resistance and can stop a transfer if large changes indicate that the buffer system is depleting Multiple gels can be transferred at the same time by placing several small gels of the same thickness side by side or by stacking two gels vertically in a carefully constructed multi layered stack The electrodes are made of the best possible materials to ensure the units last a long time Unpacking Unwrap all packages carefully and compare contents with the packing list making sure all items arrived If any part is missing contact Hoefer Inc Inspect all components for damage that may have occurred while the unit was in transit
28. oeferille Inc suojelu ehk isty varusteille saattaa olla avuton T m v line suunnitellaan sis laboratoriok yt lle vain Vain lis varusteet ja osat hyv ksyiv t tai toimitti Hoeferin oheen Inc voi k ytt k ytt miselle valvoalle ja servicing t m tuote Varoittaminen Koska t m v line voi kehitt riitt v j nnitteen ja virran tuottaa kuolettavan j rkytyksen huolta t ytyy harjoittaa toiminnossaan T m v line suunnitellaan EN61010 1 2001 s hk turvallisuusstandardin mukaisesti Silti pit isi k yt t vain ohi oikeasti koulutetut k ytt j t Lue t m kokonainen manuaalinen ennen v linett ja k ytt vain ohjeiden mukaan V linett t ytyy k ytt aina valtanuoran maalyijyst perusti oikein maadoittaa s hk verkkoaukossa K ytt vain undamaged s hk metallilankaa ja varus teita t sm llinen j nnitteille k ytt Kaikki varusteet yhdistetty korkeaan j nnitteeseen pit isi olla EN61010 1 2001IN mukaisesti Pit v lineen yht kuiva ja puhdas kuin mahdollinen Pyyhi s nn llisesti pehme ll kostealla kankaalla Anna v line kuivua t ysin ennen k ytt Ei k yt v linett extreme ilmankosteudessa 80 n yl puolella V lt tiivistymist antamalla yksik n eguili brate ymp r iv n l mp tilaan kun ottaminen v line kylmempi l mpim mp n ymp rist n Sallia riitt v n j hdytt minen varmistaa ett v lineen
29. ondensac o deixando o equili brate de unidade a temperatura ambiental quando tomar o instrumento de um mais frio a um ambiente mais quente Permitir esfriar suficiente assegura que as aberturas do instrumento n o s o cobertas Informaci n Importante Spanish Si este equipo es utilizado en una manera no especifi cado por Hoefer S a la protecci n proporcionado por el equipo puede ser da ada Este instrumento es dise ado para el uso interior del laboratorio s lo S lo accesorios y partes aprobaron o suministraron por Hoefer S a puede ser utilizado para Operar para mantener y para atender a este producto Advirtiendo Porque este instrumento puede desar rollar voltaje y corriente suficientes para producir un golpe mortal el cuidado debe ser ejercitado en su operaci n Este instrumento es dise ado de acuerdo con el EN61010 1 2001 est ndar el ctrico de seguridad No obstante debe ser utilizado s lo por operarios adecuadamente capacitados Lea este manual entero antes de utilizar el instrumento y el uso s lo seg n las instrucciones El instrumento siempre debe ser utilizado con el plomo de la tierra del cable de alimentaci n moli correcta mente a la tierra en la salida de red Utiliza alambre y equipo el ctricos s lo ilesos espec ficos para los voltajes que usted utilizar Todo equipo conectado al voltaje alto debe ser de acuerdo con EN61010 1 2001 Mantiene el instrumento tan
30. s GM PV45 1515 30 cm x 3 m 1 roll GM PV45 Hoefer Electrophoresis Reagents Tris 1 kg GR132 1 Glycine 1 kg GR125 1 SDS 500 g GR126 500 Tween 20 500 ml GR128 500 DTT 5g GR122 5 Glycerol 1 L GR124 1 Bromophenol Blue BPB 10 g GR120 10 Protein Determination Reagent 500 standard assays GR133 500 Coomassie Brilliant Blue G 250 25 g GR134 25 Coomassie Brilliant Blue R 250 25 g GR135 25 e p22 Hoefer Inc 84 October Hill Road Holliston MA 01746 Toll Free 1 800 227 4750 Phone 1 508 893 8999 Fax 1 508 893 0176 E mail support hoeferinc com Web www hoeferinc com Chroma Hoefer and MiniVE are trademarks of Hoefer Inc Contrad 70 is a trademark of Decon Laboratories Tween is a trademark of ICI Americas Inc Coomassie is a trademark of ICI plc 2012 Hoefer Inc All rights reserved Printed in the USA Hoefer
31. seco y limpio como posible Enjugue regularmente con un suave el trapo h medo Permita que el instrumento seque completa mente antes de uso No opera el instrumento en la humedad extrema encima de 80 Evite condensaci n permitiendo la unidad equilibra a la temperatura ambiente al tomar el instrumento de un m s fr o a un ambiente m s tibio Permitir refrigeraci n suficiente asegure que las aberturas del Viktig Information Swedish om denna utrustning anv nds i ett s tt som inte har specificeras av Hoefer Inc skyddet tillhandah ll vid utrustningen kan skadas Detta instrument formges f r inomhuslaboratorium anv ndning bara Bara medhj lpare och delar godk nde eller levererade vid Hoefer Inc kan anv ndas f r fungera underh lla och servicing denna produkt varna D rf r att detta instrument kan utveckla tillr ck lig sp nning och str m att producera en d dlig st t m ste vas omsorg i dess funktion Detta instrument formges i verensst mmelse med EN61010 1 2001 elektriska s kerheten standarden Icke desto mindre b r det anv ndas bara av riktigt utbildade operat rer L s denna hela handbok f re anv nda instrumentet och anv nd bara enligt under visningarna Instrumentet m ste alltid anv ndas med jorden blyet av kraften repet riktigt grounded till jorden p det huvudutloppet Anv nder bara undamaged elektrisk trad och utrust ning specifik for spanningarna
32. te elektrische Leitung und Ausr s tung spezifisch f r die Stromspannungen verwenden die Sie verwenden werden Die ganze mit der Hoch spannung verbundene Ausriistung sollte in Uberein stimmung mit EN61010 1 2001 sein Das Instrument ebenso trocken halten und reinigen wie m glich Wischen Sie regelm ig mit einem weichen befeuchten Sie Stoff Lassen Sie das Instru ment trocken v llig vor dem Gebrauch Das Instrument in der u ersten Feuchtigkeit ber 80 nicht bedienen Vermeiden Sie Kondensation die Einheit equilibrate zur Umgebungstemperatur laBend wenn Sie das Instrument von einem k lteren bis eine w rmere Umgebung nehmen Um das gen gend Abk hlen zu erlauben stellen Sie sicher dass die Offnungen des Instrumentes nicht bedeckt werden Informazioni Importanti Italiano Se quest apparecchiatura usata in un modo specificato da Hoefer Inc la protezione fornito dall apparecchiatura potrebbe essere indebolita Questo strumento disegnato per l uso di laboratorio interno solo Solo gli accessori e le parti hanno approvato o hanno fornito da Hoefer Inc potrebbe essere usato per oper are per mantenere e per revisionare questo prodotto Avvertendo Perch questo strumento pu sviluppare il voltaggio sufficiente e la corrente di produrre una scossa letale la cura deve essere esercitata nella sua operazione Questo strumento disegnato conforme mente all EN61010 1 2001 la norma d
33. to start the transfer Variable Units Range Increment Current milliamps 1 500 1 mA Time hours minutes 5 min 4 00 hours 5 min Voltage volts 0 30 read only 1 volt display mode cycles LED display 0 Aoier 3 D N o r o L a Red LED LO az 10 I LJ U LJ Dry error message During the transfer TE70XP and the TE77XP A red LED will appear in the lower right corner of the Display when voltage is applied to the transfer stack Both the Time and the Current can be changed as the transfer progresses Select mA or time using the DISPLAY MODE button and the UP and DOWN to change the value While the value is being changed the LED will blink After about 10 seconds the LED will stop flashing and show the real time run parameters Buffer Depletion One of the more common failure modes for a Western transfer is buffer depletion Buffer depletion leads to changes in pH and overheating both of which are detrimental to the transfer The TE70XP and the TE77XP instruments monitor the transfer stack resistance Large changes in this resistance indicate the depletion of the buffer system The instrument can stop a transfer before these changes lead to further problems like burning of the transfer stack If this condition happens the error message dry will be on the display The instrument has stopped the transfer If desired the cover can be removed
34. tting paper the same size as the gel or slightly smaller Gauge the thickness or number of blotting paper layers according to the amount of buffer reguired up to 300 ml of buffer may be reguired for larger gels or for transfers of 60 minutes in order to prevent the stack from drying out or the buffer from becoming depleted Saturate at least 3 pieces of the blotting paper with transfer buffer One by one center each sheet on the lower electrode and remove all trapped air by rolling a clean pipet or roller from the center toward the edges Prepare the membrane For each gel cut 1 membrane the same size as the gel or slightly smaller A larger membrane may contact an electrode panel creating a pathway by which current can bypass the gel Pre wet nitrocellulose or nylon membranes with distilled water Pre wet PVDF or other hydrophobic membranes with methanol Then soak all membrane types in transfer buffer for 2 5 minutes a O Important Stack each layer with Complete the stack care with edges parallel As each a Place the pre wet membrane onto the stack of layer is added remove all air blotting paper pockets by rolling a clean pipet from the center to the edges Add b Place the gel on the membrane a few drops of butfer to trouble Note Proteins bind to the membrane as soon as areas to help remove air pockets contact occurs so it is important to place the gel Note If not using Hoefer blotting 01000
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