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Summary of Contents for Hoefer TE 70
- Page 1 TE70 and TE77 Hoefer TE 70 TE 77 semi-dry transfer units TE70&77-IM/Rev. H0/06-04...
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Page 3: Table Of Contents
Page finder Semi-dry Transfer Unit function and description .......1 Unpacking ..........2 Specifications ..........2 Important information.......3 Operating instructions ........4 Care and maintenance........10 Troubleshooting..........11 Electrotransfer notes........13 Bibliography ..........15 Ordering information ........16 •... - 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 Ein Blitzsymbol in einem gleichseitigen Dreieck soll den Benutzer auf die Gefahr anliegender Hochspannungen hinweisen. 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...
- Page 6 Il simbolo del fulmine all’interno di un triangolo equilatero indica all’utente la presenza di un rischio di esposizione ad alte tensioni. 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...
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Page 7: Semi-Dry Transfer Unit Function And Description
Joule heat- ing. Most transfers are complete in one hour or less. The smaller TE 70 unit transfer surface is 14 × 16 cm, and the larger TE 77 transfer surface is 21 × 26 cm. -
Page 8: Unpacking
If any part is missing, contact instrument when it is: Hoefer, Inc. Inspect all components for damage • used in laboratory locations. that may have occurred while the unit was in • used as delivered from transit. -
Page 9: 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. Informations importantes Français •... -
Page 10: Operating Instructions
Operating instructions To transfer proteins, prepare the unit and trans- Note: See Electrotransfer fer stack components, assemble the stack, and notes on page 11 for transfer tips as well as two buffer reci- connect the leads to a power supply. Then run the pes and references. - Page 11 Gauge the thickness or number of blotting paper layers according to the amount of buffer required; up to 250 ml of buffer may be required for larger gels or for transfers of 60 minutes in order to prevent the stack from drying out or the buffer from becoming depleted.
- Page 12 Complete the stack a) Place the pre-wet membrane onto the stack of blotting paper. b) Place the gel on the membrane. Note: Proteins bind to the membrane as soon as contact occurs, so it is important to place the gel correctly on the first try.
- Page 13 Blotting Fig 3. Transfer stacks for gels paper, (2–3 placed side-by-side. sheets) Check electrical continuity within each stack: several sheets of buffer-soaked blotting paper must cover each gel, and each Membrane stack must fully contact both electrodes. Blotting paper, (2–3 Note: There must be no electrical sheets) contact between the two stacks.
- Page 14 If possible, set the voltage limit before connecting the transfer. transfer unit. Use only an approved power supply such as the Hoefer PS2A200. The power supply should be switched off and both the Note: Rewet dried membranes...
- Page 15 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.
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Page 16: Care And Maintenance
After the transfer is complete Turn off the power supply and disconnect the leads from the power supply. Unplug the safety-interlock lead between the cover and the base of the unit. Remove the cover slowly because the stack may adhere to it. ... -
Page 17: Troubleshooting
Troubleshooting problem solution Incomplete transfer Blank or faint Remove trapped air pockets between the gel and membrane during stack assembly. areas on the membrane Reduce the current setting to minimize the amount of gas produced by the electrolysis of buffer. Use buffer with a lower ionic strength. - Page 18 Smeared or diffuse If equilibrating before the transfer, shorten or eliminate the equilibra- tion time and/or equilibrate under coldroom conditions. band patterns If the transfer buffer contains ≥10% methanol, equilibrate the gel in transfer buffer for 30 minutes to allow it to shrink before assembling the stack.
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Page 19: Electrotransfer Notes
Electrotransfer notes • Run the transfer as soon as possible after electro- phoresis to minimize protein diffusion within the gel. • Stacked gels must all be the same size. • Limit transfers to two hours or less, rewetting the stack after 1 hour. •... - Page 20 CAPS buffer, 1X (10 mM CAPS, pH 11.0, 1 liter) CAPS (FW 221.3) 10 mM 2.2 g [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. • For a 3-buffer system, refer to Kyhse-Anderson, J. (1984).
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Page 21: Bibliography
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 Immuno- detection. In Current Protocols in Molecular Biology. 10.8.1–10.8.17. Greene Publishing and Wiley-Interscience, NY (1993). -
Page 22: Ordering Information
Ordering information product quantity code number TE 70 Semi-dry Transfer Unit, 14 × 16 cm TE70 Includes 25 sheets of blotter paper, 50 sheets of cellophane, 2 uncut masks, precut mylar mask for 7 × 8 cm gels and for 14 × 16 cm gels. - Page 23 •...
- Page 24 Noryl is a registered trademark of General Electric Company. Conrad 70, Decon and Neutrad are trademarks of Decon Laboratories, Inc. Printed in the USA Hoefer, Inc. 953 Indiana Street San Francisco, CA 94107 USA www.hoeferinc.com •...
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