Hoefer TE 22 User Manual
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Hoefer TE 22 User Manual

Tank transfer unit
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TE 22
Hoefer
TE 22
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tank transfer unit
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TE22-IM/Rev. E0/08-04

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Summary of Contents for Hoefer TE 22

  • Page 1 TE 22 Hoefer TE 22 ™ tank transfer unit TE22-IM/Rev. E0/08-04...

  • Page 3: Table Of Contents

    Page finder Transfer Electrophoresis Unit function and description ..... Specifications ......Important information .
  • Page 4 Hoefer, Inc. Hoefer, Inc. shall in no event be liable for incidental or conse- quential 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 and incorrect use of the product.
  • Page 5 Hoefer, Inc. se réserve le droit d’effectuer des modifications de ces spécifications sans aucun préavis. Garantie et responsabilité Hoefer, Inc. garantit à l’utilisateur que le produit livré a subi avec succès tous les essais prévus pour s’assurer qu’il est conforme aux spécifications et normes en vigueur. La garantie incluse dans les conditions de livraison n’est valable que si le...
  • Page 6 Hoefer, Inc. 953 Indiana Street San Francisco, CA 94107 USA support@hoeferinc.com Hoefer, Inc. behält sich das Recht vor, die Spezifikationen ohne vorhergehende Ankündigung zu ändern. Gewährleistung and haftung Hoefer, Inc. garantiert, daß das gelieferte Produkt sorgfältig auf die Einhaltung der veröffentlichten Spezifikationen getestet wurde.
  • Page 7 Hoefer, Inc. 953 Indiana Street San Francisco, CA 94107 USA support@hoeferinc.com Hoefer, Inc. se reserva el derecho a modificar las especifica- ciones sin previo aviso. Garantía y responsabilidad Hoefer, Inc. garantiza que el producto entregado ha sido probado a fondo para comprobar el cumplimiento de las espe- cificaciones publicadas.
  • Page 8 Hoefer, Inc. 953 Indiana Street San Francisco, CA 94107 USA support@hoeferinc.com Hoefer, Inc. si riserva il diritto di apportare modifiche ai dati tecnici senza preavviso. Garanzia e responsabilitá Hoefer, Inc. garantisce che prima della consegna il prodotto è stato collaudato a fondo per soddisfare i requisiti specificati.

  • Page 9: Transfer Electrophoresis Unit Function And Description

    If any part is missing, contact Hoefer, Inc.. Inspect all components for damage that may have occurred while the unit was in transit. If any part appears damaged, contact the carrier immediately.

  • Page 10: Specifications

    500 mA tions, Max. temperature 45 °C • used as delivered from Buffer required 1.5 liters, depending on Hoefer, Inc. except for the number of cassettes alterations described in in place the User Manual, and Environmental Indoor use: 4–40 °C •...
  • Page 11 Fig 1. Tank transfer unit main components. color-coded leads A power supply capable of deliver- ing up to 100 V [ and 400 to 500 mA is required. cover transfer tank. up to four cassettes fit into the slots. electrode panels (2) electrode retaining screw (2) fill...

  • Page 12: Important Information

    • If this equipment is used in a manner not specified by the manufacturer, the protection provided by the equipment may be impaired. • Only accessories and parts approved or supplied by Hoefer, Inc. may be used for operating, maintaining, and servicing this product.
  • Page 13 équipent cet appareil peuvent être rendues inéfficaces. • Seulement les accessoires et piéces detachées approuvés ou fournis par Hoefer, Inc. sont recommandés pour l’utilisation, l’entretien et réparation de cet appareil. *L'utilisation de méthanol à une concentration finale de 20% dans les tampons de transfert constitue la seule exception.

  • Page 14: Operating Instructions

    Operating instructions Perform the transfer as soon as possible after electrophoresis to minimize band diffusion. Each step is described below. Prepare the buffer Prepare a minimum of 1.5 liters of the appropri- Note: Refer to the Electrotransfer Notes section for a discussion of ate transfer buffer.
  • Page 15 ends of each length of tubing to the circulator bath ports; one to the inlet and the other to the outlet. Secure the connections with the hose clamps. Place (do not drop) a magnetic stirring bar in the buffer tank. (Dropping objects into the tank may crack the alumina plate.) Set the unit onto a magnetic stirrer and fill transfer buffer to the “Start fill level”...
  • Page 16 sponge, and then place the membrane on the blot- ting paper. Place the gel—which contains a sample that has been electrophoretically separated and equilibrated (if required) with transfer buffer—on the membrane. Gently roll a glass pipet or test tube over the gel to expel trapped air between the membrane and gel.
  • Page 17 Install the cassette(s) If transferring only one or two gels, choose the cassette positions nearest the center. The cassettes must be oriented so that the hinge side is facing up and all black panels of the cassettes are facing the same side of the transfer unit.
  • Page 18 Use only an approved power supply such as the proteins migrate toward the anode in the Towbin Tris/glycine/metha- Hoefer PS 2A200. Make sure the power supply is off nol buffer system (independent of and all controls are set to zero. Plug the color-coded...
  • Page 19 Set the power supply Constant current mode is recommended. If constant voltage mode is selected, carefully monitor the current (increased current increases Joule heating). If the current exceeds 0.4 A, decrease the voltage. If available, set the power supply timer Most transfers are complete within one hour, but larger molecules or thicker gels may require longer transfer times;...

  • Page 20: Care And Maintenance

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

  • Page 21: 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 22 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 23: 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 24 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 25 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 26 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 27 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 28 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 29 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 30: Bibliography And References

    Bibliography and references 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 31 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 32: Ordering Information

    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 QFX3/8 Blotter paper Blotter paper, sheets, 9 × 10.5 cm TE26 Companion products Hoefer PS 2A200 Power Supply, 200 V, 2A PS2A200 •...
  • Page 34 Contrad 70 and Decon 90 are trademarks of Decon Lab. Printed in the USA Hoefer, Inc. 953 Indiana Street San Francisco, CA 94107 USA www.hoeferinc.com •...

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