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6. Bibliography

Bjerrum, O.J., Larsen, K., and Heegaard, N.,

CRC

Handbook of Immunoblotting of Proteins

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

. 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

, 384–387 (1987).

Содержание TE70X

Страница 1: ...user manual um TE70X IM Rev C0 07 12 TE70X Semi dry Blotters TE70X Semi dry transfer unit TE77X Semi dry transfer unit TE70XP Semi dry transfer unit TE77XP Semi dry transfer unit ...

Страница 2: ...ent WEEE vii 1 TE70X Semi dry Blotters Description 1 Unpacking 2 Specifications 3 2 Operating instructions 4 Electrotransfer 9 3 Care and maintenance 13 4 Troubleshooting 14 5 Electrotransfer notes 17 6 Bibliography 19 7 Customer service information 20 8 Ordering information 21 ...

Страница 3: ...ána v jeho provoz Tento nástroj je určen v souladu s EN61010 1 2001 elektrické bezpečnostní normy Přesto že by měly být použity pouze řádně vyškolený operátorů Číst celé toto ruční před použitím nástroje a použití pouze v souladu s pokyny Přístroj musí být vždy používají se na výkonu zemi vést šňůra správně zemněny k zemi na síti výústce Využití pouze nepoškozené elektrické dráty a vybavení pro na...

Страница 4: ...ngen van het instrument niet bedekt zijn Tärkeää Tietoa Finnish Jos tätä varusteita käytetään tavassa ei määritetty Hoeferille Inc suojelu ehkäisty varusteille saattaa olla avuton Tämä väline suunnitellaan sisälaboratoriokäytölle vain Vain lisävarusteet ja osat hyväksyivät tai toimitti Hoeferin oheen Inc ää voi käyttää käyttämiselle valvoalle ja servicing tämä tuote Varoittaminen Koska tämä väline...

Страница 5: ...de am Hauptausgang verwendet werden Nur unbeschädigte elektrische Leitung und Ausrüs tung spezifisch für die Stromspannungen verwenden die Sie verwenden werden Die ganze mit der Hoch spannung verbundene Ausrüstung sollte in Überein stimmung mit EN61010 1 2001 sein Das Instrument ebenso trocken halten und reinigen wie möglich Wischen Sie regelmäßig mit einem weichen befeuchten Sie Stoff Lassen Sie ...

Страница 6: ... urządzenie może zostać obniżony Instrument ten jest przeznaczony do użytku w labora toriach kryty tylko Tylko akcesoriów i części zatwierdzone lub dostarczone przez Hoefer Inc mogą być wykorzystane do eksp loatacji utrzymania i obsługi tego produktu Uwaga Ponieważ ten akt prawny może być rozwinięcie odpowiednich napięcie i bieżących do wyprodu kowania śmiertelnego szoku opiekę musi być wykony wan...

Страница 7: ...te a la tierra en la salida de red Utiliza alambre y equipo eléctricos sólo ilesos especí ficos para los voltajes que usted utilizará Todo equipo conectado al voltaje alto debe ser de acuerdo con EN61010 1 2001 Mantiene el instrumento tan seco y limpio como posible Enjugue regularmente con un suave el trapo húmedo Permita que el instrumento seque completa mente antes de uso No opera el instrumento...

Страница 8: ...en Sie Kontakt mit einem autorisierten Beauftragten des Herstellers auf um Informationen hinsichtlich der Entsorgung Ihres Gerätes 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 modalità di smantella...

Страница 9: ...uits 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 c...

Страница 10: ...s arrived 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 Be sure to keep all packing material for damage claims or to use should it become necessary to return the unit ...

Страница 11: ... 0 mm Environmental Indoor use 4 40 C operating conditions Humidity up to 80 Altitude up to 2000 m Installation category II Pollution degree 2 Dimensions w d h 38 46 9 cm 15 18 3 5 in Weight Shipping 6 8 kg Unit 3 7 kg Certification EN61010 1 EN 61326 CE UL61010 1 2004 CSA 22 2 61010 1 04 p3 This declaration of conformity is only valid for the instrument when it is used in laboratory locations use...

Страница 12: ... 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...

Страница 13: ...ean pipet or roller from the center toward the edges 5 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 typ...

Страница 14: ...ith care with edges parallel As each layer is added remove all air pockets by rolling a clean pipet from the center to the edges Add a few drops of buffer to trouble areas to help remove air pockets Note If not using Hoefer blotting paper additional pieces may be required to achieve desired results Mask opening approx 2 mm smaller than the gel on all sides Optional Blotting paper 2 3 sheets Blotti...

Страница 15: ...nuity 7 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 Mask optional Blotting paper 2 3 sheets Blotting paper 2 3 sheets Gel Membrane Note When transferring multiple gels transfer efficiency depends on such factors as gel thickness gel position in the stack transfer buffer membrane type and ...

Страница 16: ... paper 2 3 sheets Blotting paper 1 sheet Cellophane Mask optional Gel Membrane Blotting paper 2 3 sheets Gel Membrane Fig 4 Transfer stack for stacked gels Separate the stack for each gel with a sheet of cellophane ...

Страница 17: ...eed 0 8 mA cm2 of the gel surface or 500 mA maximum If transferring several layers of gels a longer time may be required 3 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 emp...

Страница 18: ...The UP and DOWN arrows to change the value for the current and time 2 Press the START STOP button 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 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...

Страница 19: ... 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 con...

Страница 20: ...here to it 2 Remove and dispose of the upper blot papers 3 Remove the gel s Optional Stain gel to check for residual protein left in the gel 4 If desired label the gel contact side of the membrane with a soft pencil 5 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 6 Remove the remaining blot ...

Страница 21: ...washer 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 ...

Страница 22: ... 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 methan...

Страница 23: ...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 ...

Страница 24: ...ng 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 Maximum voltage 30 V has been reached lower than setting Buffer may be depleted No output current Tra...

Страница 25: ...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 v v methanol pH 8 3 1 liter Tris FW 121 1 25 mM 3 0 g Glycine FW 75 07 192 mM 14 4 g SDS FW 288 4 0 1 3 5 mM 1 0 g 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...

Страница 26: ...ulfonic 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 Transfer efficiency varies depending on the gel concentration which can be optimized For more information refer to Smejkal and Gallagher 1994 ...

Страница 27: ...ins 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 10 7 1 10 7 3 Greene Publish ing and Wiley Interscience N...

Страница 28: ...erinc 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 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 pr...

Страница 29: ...70XP Includes 25 sheets of blotting paper 50 sheets of cellophane and 2 solid masks TE77XP Semi dry Transfer Unit 21 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 18 5 cm 4 TE74 Porous cellophane 20 35 5 cm 50 TE73 Blotting paper precut 14 16 cm 25 TE76 1416 TE77X and TE77XP Solid masks 23 27 5 cm 4 TE78 ...

Страница 30: ...2320 30 cm 3 m 1 roll GM NC45 0 2 µm pore size 30 cm 3 m 1 roll GM NC22 PVDF membrane 0 45 µm pore size 10 10 cm 10 sheets GM PV45 1010 15 15 cm 5 sheets GM PV45 1515 30 cm 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 5 g GR122 5 Glycerol 1 L GR124 1 Bromophenol Blue BPB 10 g GR120 10 Protein Determinati...

Страница 31: ...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 ...

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