Hoefer TE42 User Manual
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Hoefer TE42 User Manual

Transfer electrophoresis units
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Hoefer TE42 and TE62
Transfer electrophoresis units
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TE42-IM

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Summary of Contents for Hoefer TE42

  • Page 1 Hoefer TE42 and TE62 Transfer electrophoresis units TE42-IM...

  • Page 2: Table Of Contents

    Contents Important Information ..........ii Waste Electrical and Electronic Equipment (WEEE) .......vii Transfer Electrophoresis Unit function and description ........1 Specifications ............2 Operating instructions ..........4 Care and maintenance .........12 Mains power module ..........13 Troubleshooting ...........15 Electrotransfer notes ..........17 Bibliography ............24 Ordering information ..........26 •...

  • Page 3: Important Information

    Important Information – English • Rozeslat pouze voda nebo 50/50 voda/ethyleng- lykolu prostřednictvím výměník tepla je li to vybav- • If this equipment is used in a manner not speci- ena. Nemají připojení výměník tepla s vodními fied by Hoefer, Inc. the protection provided by the setřepná nebo jakékoli chladicí kapaliny zdroje, kde equipment may be impaired. tlak vody je neregulo. • This instrument is designed for indoor laboratory • Nikdy zavést prostředek proti zamrznutí nebo use only. jakákoli organická rozpouštědla do jakékoli části z • Only accessories and parts approved or supplied tohoto nástroje. Rozpustidlům způsobí nenapravi- by Hoefer, Inc. may be used for operating, main- telné poškození jednotka! taining, and servicing this product. • Nejsou provozována s pufru teplotách nad • Only use a power supply that is CE marked or maximální stanovenou technickými specifika- safety certified by a nationally recognized testing cemi. Přehřátí způsobí nenapravitelné poškození laboratory. jednotka! • The safety lid must be in place before connecting Vigtig Information – Danish the power supply leads to a power supply.
  • Page 4 Belangrijke Informatie – Dutch on kansallisesti tunnustettnut testaaminen labo- ratoriota. • Indien deze uitrusting in een manier wordt • Turvallisuuskansi täytyy olla paikallaan ennen gebruikt die niet door Hoefer, Inc. is gespecificeerd yhdistäminen käyttöjännitelyijyjä käyttöjännit- de bescherming die door de uitrusting is verzorgd teeseen. kan worden geschaad. • Kiertää kaikki käyttöjännitevalvonnat ja irrottaa • Dit instrument is voor binnenlaboratoriumgebruik valtalyijyt ennen poistaminen turvallisuuskantta. enkel ontworpen. • Kiertää vain vesi tai 50/50 vesi/ethyleneä glycol • Enkel onderdelen en delen keurden goed of siinä tapauksessa varustetun lämmönvaihtimen leverden door Hoefer, Inc. kan voor het bedienen läpi. Älä yhdistä lämmönvaihdinta vesinapautuk- worden gebruikt, handhavend en onderhouden seen eikä jäähdytysnestelähteeseen, missä vesi- van dit product. paine on unregulated. • gebruik Enkel een netvoeding die CE is markeerde • Pakkasneste eikä orgaaninen liuotin välineen of veiligheid die door een is gecertificeerd die osassa ei esitele Koskaan. Orgaaniset liuottimet nationaal is herkend testene laboratorium. aiheuttavat korvaamattoman vahingon yksikköön! • Het veiligheidsdeksel moet in plaats voor het...
  • Page 5 Informazioni Importanti – Italian l’instrument. Les dissolvants organiques causeront des dommages irréparables à l’unité! • Se quest’apparecchiatura è usata in un modo • Ne pas fonctionner avec les températures de specificato da Hoefer, Inc. la protezione fornito tampon au-dessus du maximum a spécifié des dall’apparecchiatura potrebbe essere indebolita. spécifications techniques. La surchauffe causera • Questo strumento è disegnato per l’uso di labora- des dommages irréparables à l’unité ! torio interno solo. • Solo gli accessori e le parti hanno approvato o Wichtige Informationen – German hanno fornito da Hoefer, Inc. potrebbe essere • Wenn diese Ausrüstung gewissermaßen nicht usato per operare, per mantenere, e per revisionare angegeben durch Hoefer, Inc. verwendet wird, questo prodotto. kann der durch die Ausrüstung zur Verfügung • usa Solo un alimentatore che è CE ha marcato o la gestellte Schutz verschlechtert werden. sicurezza certificato da un nazionalmente ricon- • Dieses Instrument wird für den Innenlaborge- osciuto testando il laboratorio. brauch nur dafür entworfen. • Il coperchio di sicurezza deve essere nel luogo • Nur Zusätze und Teile genehmigten oder lieferten...
  • Page 6 • Nie działają w buforze temperatury powyżej kraftforsyningene blyene til en kraftforsyning. maksymalnego określone specyfikacje techniczne. Przegrzania spowoduje nieodwracalne szkody dla • Vender all kraftforsyningsstyring av og frakopler jednostki! kreftene blyene før fjerning sikkerheten lokket. • Sirkulerer bare vann eller 50/50 vann/ethylene Informações Importantes – glykol gjennom oppvarmingen veksleren i så fall Portuguese utstyrer. Ikke forbind oppvarmingen veksleren til en vanntapp eller noe kjølemiddelkilde hvor • Se este equipamento é usado numa maneira não vannet trykket er unregulated. especificada por Hoefer, Inc. que a protecção forne- • Introduserer Aldri antifreeze eller noe organisk cida pelo equipamento pode ser comprometida. løsemiddel inn i noe del av instrumentet. Organi- • Este instrumento é projectado para uso de interior ske løsemiddler vil forårsake irreparabel skade på de laboratório só. enheten ! • Só acessórios e partes aprovaram ou forneceu por • Driver med buffertemperaturer over maksimum Hoefer, Inc. pode ser usada para operar, manter, e ikke spesifiserte teknisk spesifikasjoner. Å overop- servicing este produto. pheting vil forårsake irreparabel skade på enheten ! • Só usa um estoque de poder que é CE marcou ou segurança registrada por um nacionalmente recon-...
  • Page 7 • Inför aldrig kylvätska eller något organiska • Sólo utiliza una alimentación que es CE marcó o lösningsmedel in i någon del av instrumentet. la seguridad certificada por un nacionalmente Organiskt lösningsmedel ska orsaka irreparable reconocido probando el laboratorio. skada till enheten! • La tapa de la seguridad debe estar en el lugar • Använd inte med buffert temperaturer över antes de conectar la alimentación lleva a una det högsta angivna tekniska specifikationerna. alimentación. Överhettning skulle orsaka irreparabla skador på enheten! • Apaga todos controles de alimentación y desco- necta los plomos del poder antes de quitar la tapa de la seguridad. • Circula sólo agua o 50/50 glicol de agua/etileno por el intercambiador de calor si ése es el caso equiparon. No conecte el intercambiador de calor a un toque de la agua ni cualquier fuente del líquido refrigerante donde la presión del agua está libre. • Nunca introduce anticongelante ni algún solvente orgánico en cualquier parte del instrumento. Los solventes orgánicos causarán daño irreparable a la unidad! • No opera con temperaturas de búfer encima del máximo especificó especificaciones técnicas. Reca- lentar causará daño irreparable a la unidad! Viktig Information –...

  • Page 8: Waste Electrical And Electronic Equipment (Weee)

    Waste Electrical and Electronic Equipment (WEEE) English This symbol indicates that the waste of electrical and electronic equipment 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 equipment.

  • Page 9: Transfer Electrophoresis Unit Function And Description

    The TE62 contains a heat exchanger in the base. Buffer is separated from the coolant by a heat-conducting alumina plate. The TE42 is not equipped with a buffer cooling system. If cooling is required, an immersible heat exchanger can be ordered separately.

  • Page 10: Specifications

    Altitude up to 2000 m Installation category II Pollution degree 2 Dimensions (w × d × h) TE42: 28 × 13 × 30.5 cm (11 × 5.1 × 12 in.) TE62: 28 × 16.5 × 32 cm (11 × 6.5 × 12.5 in.) Product certifications EN61010–1, UL3101–1,...
  • Page 11 Foam sponges, 3 mm thick (8) Blotting paper, sheets (25) color-coded electrode panels (2) Tank fill levels Note: An immersible heat TE62: coolant safety cassette hook exchanger (Code no. valve and coolant and holder TE47) can be ordered ports (2) separately for the TE42. •...

  • Page 12: Operating Instructions

    Operating instructions Perform the transfer as soon as possible after electrophoresis to minimize band sample diffu- sion. Each step is described below. Prepare the buffer Prepare a minimum of 5 liters of the appropriate Note: Refer to the Electrotransfer Notes section for a discussion of transfer buffer.
  • Page 13 TE42 Lower the heat exchanger (ordered separately, or use the heat exchanger supplied with the Hoefer SE600 Note: The relief valve opens if the Gel Electrophoresis Unit if you have one) into the pressure in the heat exchanger exceeds 0.7 bar (10 psi) above lower chamber, fitting the ports into the notches in atmospheric pressure.
  • Page 14 Assemble the transfer cassette Note: Always wear gloves when Pre-wet nitrocellulose or nylon membranes with handling membranes to avoid distilled water. Pre-wet PVDF or other hydrophobic getting fingerprints on them. membranes in methanol. Then soak all membrane Important!  Take great care in types in transfer buffer for 2–5 minutes.
  • Page 15 Close the cassette and press lightly to lock the tabs. The assembled cassette should hold the gel in firm contact with the membrane without squeezing the gel. If the stack seems loose, add sheets of blot- ting paper; if the stack seems tight, replace the top sponge (above the gel) with a sheet of blotting paper.
  • Page 16 (The submersible heat exchanger, if used in the TE42, fills the two center slots, so only two cassettes can be placed in the outside slots.) The cassettes must be oriented so that the hinges face up and so that the black side of each cassette faces the black cathode panel.
  • Page 17 Transfer Take care in orienting all system components so that the electric field applied causes all species to migrate toward the membrane. The migration direction depends on both the characteristics of the sample and the pH of the transfer buffer. If the species of interest is negatively charged in the transfer buffer and the stack is assembled so that the membrane is nearest the grey side...
  • Page 18 Note: The two red caps on the lid Use only an approved power supply such as the accommodate the banana plugs on Hoefer PS2A200, PS200HC, or PS300B. Make sure the SE600 model immersible heat the power supply is off and all controls are set to exchanger (irrespective of zero.
  • Page 19 After the transfer is complete Note: It is a good idea to stain the Turn the voltage and current settings to zero and turn gel to determine the completeness off the power supply. Disconnect the leads from the of the transfer. power supply jacks.

  • Page 20: Care And Maintenance

    Care and maintenance Cleaning • Do not autoclave or heat any part above 45 °C. * Use ≤20% methanol (methyl • Do not expose to alcohols or organic solvents!* alcohol) in transfer buffers is the only exception. • Never use abrasive detergents. •...

  • Page 21: Mains Power Module

    Mains power module Fig 3. The mains power module is insert screw- driver in this located on the back panel. notch to open the cover. Important!  Fuses protect equip- ment by disconnecting loads too large for the instrument’s circuit design, so it is imperative that insert the screw- fuses are replaced only with fuses driver blade behind...
  • Page 22 Pull the fuse out of its cassette and inspect. If the fuse element is burned or broken, replace the fuse with an identical type. If the fuse appears to be intact, check it with a multi-meter. (A reading of 1Ω or less indicates the fuse is still usable.) After placing a good fuse into the cassette, slide it into the power module, making sure the arrow on the...

  • Page 23: Troubleshooting

    Troubleshooting problem solution Incomplete transfer Blank areas on Remove all trapped air pockets in the transfer stack assembly: the membrane assemble the stack while it is submerged in transfer buffer, gently press on each sponge as it is added to the stack, and roll a glass pipette or test tube over the membrane and gel to eliminate all air bubbles.
  • Page 24 problem solution Diffuse band patterns Transfer immediately after electrophoretic separation. If equilibrat- ing before the transfer, shorten or eliminate the equilibration time or move the gel to the cold room during equilibration. If transfer buffer contains methanol (≥10%), equilibrate the gel in transfer buffer for 30 minutes to allow it to shrink before assembling the stack.

  • Page 25: Electrotransfer Notes

    Electrotransfer notes Electrophoretic transfer advantages Electrophoretic transfer of proteins and nucleic acids is much faster than the blotting methods first described by Southern for DNA, Alwine et al. for RNA, or Renart et al. for proteins. The tank transfer method uses high current to reduce the transfer time of most samples to 45–60 minutes.
  • Page 26 Factors affecting the transfer Parameters such as sample characteristics, membrane type, gel pore size, and the transfer buffer used all contribute to the transferability of macromolecules, and should be kept in mind when developing a protocol. Very small molecu- lar species, for instance, migrate quickly but often do not bind as well as larger molecules;...
  • Page 27 Instrument guidelines Cooling Considerable Joule heat is generated during any transfer because of the high current employed, so active cooling is recommended, especially for transfers requiring more than one hour, protein transfers where biological activity must be retained, or transfer of nucleic acids. (The high conductivity of the phosphate buffer used by Bittner et al.
  • Page 28 Protein transfers Study summaries Gershoni and Palade (1982) investigated factors affecting protein recovery from SDS gels to nitrocellulose or DBM paper. According to their findings, methanol in the Towbin buffer system is necessary to achieve efficient binding to nitro- cellulose. Methanol improves binding in part by removing protein-bound SDS.
  • Page 29 Protein transfer buffers Use a buffer with low ionic strength, such as the two listed below, to prevent overheating. Use the alternate CAPS buffer when Tris cannot be used, as in peptide sequencing. CAPS can improve transfer because of its effect on the charge of the protein (see Matsudaira, 1987).
  • Page 30 Nucleic acid transfers Nucleic acids must normally be transferred in denatured form for most efficient binding. RNA is normally denatured with glyoxal before sepa- ration or separated in denaturing gels contain- ing formaldehyde or methyl mercury. However, double stranded DNA is usually denatured in the gel with NaOH.
  • Page 31 EDTA solution (0.5 M EDTA, pH 8.0, 100 ml) EDTA·2H O (FW 372.2) 0.5 M 18.6 g Dissolve in 70 ml distilled water. Adjust to pH 8.0 with 10 M NaOH (approx. 5 ml), then add distilled water to 100 ml. DNA transfer buffer, 10X (10X Tris-borate-EDTA (TBE) , pH ~8.2, 1 liter)

  • Page 32: Bibliography

    Bibliography Alwine, J.C., Kemp, D.J., and Stark G.R., Method for detection of specific RNAs in agarose gels by trans- fer to DBM paper and hybridization with DNA probes. Proc. Natl. Acad. Sci. USA. 74, 5350–5354 (1977). Bittner, M., Kupferer, P., and Morris, C.F., Electropho- retic transfer of proteins and nucleic acids from slab gels to diazobenzyloxymethyl cellulose or nitrocellulose sheets.
  • Page 33 Lin, W., and Kasamatsu, H., On the electrotrans- fer of polypeptides from gels to nitrocellulose membranes. Anal. Biochem. 128, 302–311 (1983). Matsudaira, P. Sequence from Picomole Quantities of Proteins Electroblotted onto Polyvinylidene Difluo- ride Membranes. J. Biol Chem. 262, 10035 (1987). Ohmsted, J.B., Affinity purification of antibodies from diazotized paper blots of heterogeneous protein samples.

  • Page 34: Ordering Information

    Sponges, Dacron, 6-mm thick. TE45 Sponges, foam, 6-mm thick. TE45F Sponges, foam, 3-mm thick. TE45F-1/8 Lid with cables for TE42 or TE62. TE49 High voltage leads SE6056-HV Quick-fit coupler body, female, to fit 9.5 mm (3/8") ID tubing QF3/8 Quick-fit coupler body, male, to fit 9.5 mm (3/8") ID tubing...
  • Page 35 Hoefer, Inc. 32 Scotland Blvd, Ste. 9, Bridgewater, MA02324, USA Toll Free: 1-800-813-0488 Phone:1-508-807-4665 E-mail: support@hoeferinc.com Web: www.hoeferinc.com Hoefer is a registered trademark of Hoefer, Inc. © 2023 Hoefer, Inc. — All rights reserved. •...

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