Hoefer TE 22 User manual

user manual
TE 22
um TE22-IM/Rev. E0/08-04
Hoefer™ TE 22
tank transfer unit

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Page finder
Transfer Electrophoresis Unit
function and description . . . . . . . . . . . . . . . . . . . . . 1
Specifications . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2
Important information . . . . . . . . . . . . . . . . . . . . . . . . 4
Operating instructions . . . . . . . . . . . . . . . . . . . . . . . . 6
Care and maintenance. . . . . . . . . . . . . . . . . . . . . . . 12
Troubleshooting . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13
Electrotransfer notes . . . . . . . . . . . . . . . . . . . . . . . . 15
Bibliography and references . . . . . . . . . . . . . . . . . . 22
Ordering information . . . . . . . . . . . . . . . . . . . . . . . . 24

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Safety warnings and precautions
Important user information
Please read this entire manual to fully understand the safe
and effective use of this product.
The exclamation mark within an equilateral triangle is
intended to alert the user to the presence of important
operating and maintenance instructions in the literature
accompanying the instrument.
The lightning symbol within an equilateral triangle is intended
to alert the user to the risk of exposure to high voltages.
Should you have any comments on this manual, we will be
pleased to receive them at:
Hoefer, Inc.
953 Indiana Street
San Francisco, CA 94107 USA
[email protected]
Hoefer, Inc. reserves the right to make changes in the specifi-
cations without prior notice.
Warranty and liability
Hoefer, Inc. guarantees that the product delivered has been
thoroughly tested to ensure that it meets its published specifi-
cations. The warranty included in the conditions of delivery is
valid only if the product has been installed and used accord-
ing to the instructions supplied by 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.
English
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Renseignements importants d’utilization
Pour une bonne compréhension et une utilisation en sécurité
maximale, il convient de lire entièrement ce manuel.
Dans la documentation qui accompagne l’instrument un point
d’exclamation dans un triangle équilatéral a pour but d’attirer
l’attention de l’utilisateur sur des instructions importantes de
fonctionnement ou de maintenance.
Le symbole de l’éclair dans un triangle équilatéral a pour
objet d’attirer l’attention de l’utilisateur sur un danger
d’exposition à la haute tension.
Tous vos commentaires sur ce manuel seront les bienvenus et
veuillez les adresser à:
Hoefer, Inc.
953 Indiana Street
San Francisco, CA 94107 USA
[email protected]
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
produit a été installé et utilisé conformément aux instructions
fournies par Hoefer, Inc.
La société Hoefer, Inc. ne sera en aucun cas responsable de
tout dommage causé directement ou indirectement par toute
utilisation incorrecte ou non approuvée du produit ou décou-
lant de cette utilisation, y compris toute perte de bénéfice
ou de recettes, toute perte de perspectives commerciales,
tout empêchement d’utilisation et tout autre risques ayant un
rapport avec l’utilisation du produit, mais sans aucune limita-
tion quant à la nature de ces dommages.
Français
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Wichtige benutzerinformationen
Für ein vollständiges Verständnis und eine sichere Hand-
habung dieses Produktes ist es notwendig, daß der Benutzer
dieses Handbuch vollständig durchliest.
Ein Ausrufezeichen in einem gleichseitigen Dreieck soll
den Benutzer auf die Anwesenheit wichtiger Betriebs- und
Wartungsanweisungen in der dem Gerät beiliegenden Doku-
mentation hinweisen.
Ein Blitzsymbol in einem gleichseitigen Dreieck soll den
Benutzer auf die Gefahr anliegender Hochspannungen
hinweisen.
Wenn Sie Anmerkungen zu diesem Handbuch haben, dann
senden Sie diese bitte an:
Hoefer, Inc.
953 Indiana Street
San Francisco, CA 94107 USA
[email protected]
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. Die in den Lieferbedingungen näher erläuter-
ten Gewährleistungsansprüche gelten nur dann, wenn das
Produkt gemäß den von Hoefer, Inc. gelieferten Anweisungen
installiert und benutzt wurde.
Hoefer, Inc. übernimmt keinerlei Haftung für Schäden oder
Folgeschäden, einschließlich, aber nicht begrenzt auf Gewin-
neinbußen, Einkommensverluste, entgangene Geschäftsab-
schlüsse, Verlust der Gebrauchsfähigkeit oder andere Verluste,
die wie auch immer durch eine fehlerhafte oder unsachgemäße
Verwendung des Produkts verursacht wurden.
Deutsch
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• pv
Información importante para el usuario
Para comprender el producto y utilizarlo con seguridad es
necesario leer este manual en su totalidad.
El signo de admiración en un triángulo equilátero en el
manual, advierte al usuario sobre la presencia de instruccio-
nes importantes de operación y mantenimiento del aparato.
El símbolo del rayo en un triángulo equilátero alerta al usuario
sobre el riesgo de exposición a altas tensiones.
Si desearan hacer algún comentario sobre este manual,
tengan la amabilidad de remitirlo a:
Hoefer, Inc.
953 Indiana Street
San Francisco, CA 94107 USA
[email protected]
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. La garantía incluida en las condicio-
nes de entrega sólo es válida si el producto se ha instalado
y utilizado de acuerdo con las instrucciones entregadas por
Hoefer, Inc.
Hoefer, Inc. no será responsable, bajo ningún concepto,
de daños directos o indirectos, incluyendo sin limitación la
pérdida de beneficios, la pérdida de ingresos, la pérdida de
oportunidades de negocio, la pérdida de utilización y otras
consecuencias relacionadas, cualquiera que sea la causa, que
se deban a la utilización defectuosa e incorrecta del producto.
Español
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Informazioni importanti per l’operatore
Per un utilizzo sicuro del prodotto, leggere attentamente
l’intero contenuto del presente manuale.
Il punto esclamativo all’interno di un triangolo equilatero
indica all’operatore la presenza di importanti istruzioni di
funzionamento e manutenzione nella documentazione allegata
al prodotto.
Il simbolo del fulmine all’interno di un triangolo equilatero
indica all’utente la presenza di un rischio di esposizione ad
alte tensioni.
Si prega di inviare eventuali commenti al presente manuale a:
Hoefer, Inc.
953 Indiana Street
San Francisco, CA 94107 USA
[email protected]
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.
La garanzia inclusa nelle condizioni di consegna risulta valida
solamente se il prodotto è stato installato ed utilizzato nel
rispetto delle istruzioni fornite da Hoefer, Inc.
Hoefer, Inc. non potrà essere ritenuta responsabile di inci-
denti o danni consequenziali, inclusi’ma non limitati’a perdite
di profitti, mancato guadagno, perdite di affari, difetti di
funzionamento e relative esposizioni, dovuti ad un utilizzo non
corretto del prodotto.
Italiano
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Transfer Electrophoresis Unit
function and description
The Hoefer™ TE22 Tank Transfer unit rapidly
transfers proteins, DNA, or RNA from up to
four small-format polyacrylamide or agarose gels
onto a membrane. Gels and membranes are held
by a cassette, which is submerged into the trans-
fer tank. Molecules migrate under an electric
field to the membrane, where they are bound.
The transfer buffer temperature can be
controlled by circulating cooled liquid through
the heat exchanger in the base. The buffer is
separated from the coolant by a heat-conducting
alumina plate.
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. 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.

• p2
Specifications
Gel size up to four 9 × 10 cm gels
Max. wattage 50 W
Max. voltage 100 V [
Max. amperage 500 mA
Max. temperature 45 °C
Buffer required 1.5 liters, depending on
the number of cassettes
in place
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 × h × d) 14 × 24 × 16.5 cm
(5.5 × 9.5 × 6.5 in)
Product certifications EN61010–1, UL3101–1,
CSA C22.2 1010.1, CE
This declaration of
conformity is only valid for
the instrument when it is:
• used in laboratory loca-
tions,
• used as delivered from
Hoefer, Inc. except for
alterations described in
the User Manual, and
• connected to other CE
labeled instruments or
products recommended or
approved by Hoefer, Inc.

• p3
Included but not shown:
• Gel cassettes (4)
• Foam sponges, 6 mm thick (4)
• Foam sponges, 3 mm thick (8)
• Blotting paper, sheets (25)
cover
electrode panels (2)
electrode retaining
screw (2)
transfer tank.
up to four
cassettes fit into
the slots.
heat exchanger
ports (2)
heat exchanger
pressure safety
valve
cassette hook
color-coded leads
fill
levels
Fig 1. Tank transfer unit main
components.
A power supply capable of deliver-
ing up to 100 V [ and 400 to
500 mA is required.

• p4
Important information
• The safety lid must be in place before connecting the power
leads to a power supply.
• Turn all power supply controls off and disconnect the power
leads before removing the safety lid.
• Rinse only the electrodes (not the banana plugs) with
distilled water before use.
• Circulate only water or 50/50 water/ethylene glycol through
the heat exchanger. Never use anti-freeze mixtures in the
heat exchanger.
• Do not operate with buffer temperature above 45 °C. All plastic
parts are rated for 45 °C continuous duty.
• Circulate coolant through the heat exchanger to minimize
heating. Overheating will cause irreparable damage to the
unit! Do not connect the heat exchanger to a water tap or
any coolant source where the water pressure is unregulated.
• For longer runs you can control heating somewhat by chill-
ing the buffer before use, running the unit in a cold room,
or both.
• When assembling the transfer cassette, use only the
required amount of gel support materials (sponges and
blotting paper) to prevent overstuffing the cassette. Excess
materials may result in cassette damage.
• Never expose any part of the instrument to alcohols or
organic solvents.* Alcohols or organic solvents will cause
irreparable damage to the unit!
• 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.
* Use of ≤20% methanol (methyl alcohol) in transfer buffers is the
only exception.
English
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• p5
Informations importantes
• Le couvercle de sécurité doit être en place avant de
brancher les prises au générateur.
• Eteindre le générateur et débrancher les prises avant
d’enlever le couvercle de sécurité.
• Rinser seulement les électrodes (pas les “banana-plugs”)
avec de l’eau distillée juste avant l’utilisation.
• Faire circuler seulement de l'eau ou bien une solution
eau/ethylène glycol dans une proportion de 50:50, dans
l'échangeur de chaleur. Ne jamais faire circuler une solution
d'antigel dans l'échangeur de chaleur.
• Ne pas utiliser avec un tampon à une température au dessus
de 45 °C. Toutes les piéces en plastique sont prévues pour
résister à une température constante de 45 °C.
• Faire circuler l’eau dans l’échangeur vertical pour minimiser
l’échauffement afin d’éviter des dommages irréparables à
l’instrument. Ne pas connecter l’échangeur vertical à circu-
lation d’eau à un robinet ou quelque source de refroidisse-
ment dont la pression n’est pas régulière.
• Pour des coulages plus long, on peut aussi contrôler la
température en refroidissant le tampon avant l’utilisation
et/ou en utilisant l’instrument dans une chambre froide.
• Utiliser uniquement la quantité prescrite d’éponges et de
papier filtre afin que la cassette ne soit pas trop pleine.
Trop de materiels peut endommager la cassette.
• Ne jamais exposer l'appareil à des alcools et des solvents
organiques.* Les alcools et les solvents organiques causent
des dommages irréparables à l'appareil.
• Si l’instrument n’est pas utilisé en conformité avec les
recommandations du fabriquant, les protections de sécurité
qui 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.
Français
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• p6
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-
ate transfer buffer. Chill the buffer before use if
possible.
Prepare the unit
1
Rinse the transfer tank and cassettes with
distilled water.
2
Active cooling is optional but strongly recommended. If
no active cooling will be used, go to step 3.
Note: Connect the heat exchanger to a circulator
bath such as the MultiTemp™ III.
Circulate only water or 50/50 water/ethylene glycol
to prevent damage to the unit.
The circulator pump must not generate a pressure
greater than 0.7 bar (10 psi) above atmospheric
pressure.
Set the temperature to 10 °C or higher if circulat-
ing only water. If using 50/50 ethylene glycol/
water, the temperature can be set lower.
Start the circulator bath at the same time as
the transfer.
First attach tubing to the red pressure relief valve
between the water inlet and outlet ports and insert
the free end into the bath or other container or drain
to catch any pressure relief overflow. The relief valve
opens if the pressure within the heat exchanger
exceeds 10 psi.
Prepare two lengths of 9 mm (3/8”) vinyl or silicone
tubing. Slide hose clamps (4 total) onto each end of
two lengths of tubing. Attach one end of each length
of tubing to a heat exchanger port. Attach the free
Note: For quick and easy connec-
tions, install Quick-fit coupler
fittings with valves in the line.
Note: Refer to the Electrotransfer
Notes section for a discussion of
membranes and buffers.

• p7
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.
3
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”
line on the front of the tank. (This requires approxi-
mately 0.7 liters.)
4
Set the stirrer to low-medium, which accomplishes
buffer circulation without forcing buffer through the
cassettes.
Note: Even if no cooling is required for your
system, the buffer should be circulated with a
stirrer to avoid buffer depletion at the electrodes.
Assemble the transfer cassette
1
Pre-wet nitrocellulose or nylon membranes with
distilled water. Pre-wet PVDF or other hydrophobic
membranes in methanol. Then soak all membrane
types in transfer buffer for 2–5 minutes.
2
Open the cassette by releasing both latch tabs along
the edge opposite the hinges. Place the opened
cassette into a tray filled with at least 3 cm of trans-
fer buffer.
3
Assemble the transfer stack so that molecules will
migrate toward the membrane. For negatively charged
macromolecules (such as nucleic acids and most
proteins), build the stack on the grey half of the
cassette (and then later position the lid so that the
grey side faces the red lead, or anode, +).
Place one 3 mm-thick foam sponge on the opened
submersed cassette and press gently until all air is
expelled. Place one sheet of blotting paper on the
Note: Always wear gloves when
handling membranes to avoid
getting fingerprints on them.
Important! Take great care in
removing all air bubbles at each
step because the presence of
air bubbles, especially between
the membrane and gel, blocks
transfer.

• p8
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. Cover the gel with a sheet of blotting paper
and then place a sponge of the proper thickness (see
the diagram below), again pressing gently to expel
trapped air.
4
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 blotting paper;
if the stack seems tight, replace the top sponge (over
the gel) with a sheet of blotting paper. If you remove
the bottom sponge (below the gel), substitute at least
two sheets of blotting paper to create space between
the membrane and the cassette panel.
one 3 mm sponge for gels
>1.5 mm
—OR—
one 6 mm sponge for gels
≤1.5 mm.
blotting paper
blotting paper
one 3 mm sponge
gel
membrane
Fig 2. Transfer stack assembly.
The stack is oriented so that
negatively charged molecules
migrate toward the grey anode, +.
Important! Do not overstuff the
cassette.
Note: Try to place the gel
correctly the first time because
proteins may begin to transfer
immediately; once transfer has
begun, moving the gel will distort
results or cause “shadow bands”
on the blot.
The cassette panels are color
coded: black (top) = cathode side
grey (bottom) = anode side
Assemble the cassette in a tray containing transfer
buffer about 3 cm deep.

• p9
Install the cassette(s)
1
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.
Work quickly when moving the assembled cassette(s)
to the tank to avoid draining the sponges: Place the
tray holding the cassette(s) near the tank, lift out one
cassette at a time, and slide it into a set of vertical
slots. Do not discard the buffer.
2
Once in place, tap the cassette lightly until most air
bubbles are dislodged. (A few small bubbles in the
sponges are unlikely to interfere with the transfer.)
3
Inspect the buffer level. Add or remove buffer as
required so that the level falls between the minimum
and maximum buffer level lines. (Buffer above the
maximum buffer level line may cause corrosion of the
electrical contacts.)

• p10
Final assembly and transfer
1
Install the lid
The cassettes are color coded to match the leads in
the lid. To transfer toward the anode, orient the lid so
that the grey half of the cassette faces the anode (+),
or red lead, and the black half of the cassette faces
the cathode (–), or black lead. Make sure the banana
plugs seat into the connectors in the lid.
2
Use only an approved power supply such as the
Hoefer PS 2A200. Make sure the power supply is off
and all controls are set to zero. Plug the color-coded
leads from the lid of the transfer unit into the power
supply — the red lead into the red output jack, and
the black lead into the black output jack. In most
systems, the red lead is the anode (+), and the black
lead is the cathode (–).
3
Cooling is strongly recommended
Any setting that results in higher than 5 W of power
will generate enough heat to require active heat
control. A refrigerated circulator bath using water
should be set to about 10 °C. (If using 50/50 ethyl-
ene glycol/water, the temperature can be set lower.)
Chill the buffer before use if possible.
Note: Take care in orienting the
lid so that all species migrate
toward the membrane when the
electric field is applied. 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 of the cassette, then
this side faces the anode (+). Most
proteins migrate toward the anode
in the Towbin Tris/glycine/metha-
nol buffer system (independent of
the presence of SDS), and under
most conditions, nucleic acids
are negatively charged and also
migrate toward the anode.
Important! Never allow the buffer
temperature to exceed 45 °C.
Excessive heat will cause the unit
to warp.
Typical transfer parameters
Parameters for your sample and buffer system
must be determined empirically.
protein nucleic acids
Buffer Towbin 1X TBE or 1X TAE
Current (A) 0.4 0.3
Voltage (V) ~100 50
Transfer time ~1 hour ~1 hour
Coolant temp. 10 °C 10 °C or less

• p11
4
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.
5
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; the optimum transfer time for each
system must be determined empirically.
After the transfer is complete
1
Turn the voltage and current settings to zero and turn
off the power supply. Disconnect the leads from the
power supply jacks.
2
Lift off the lid. Use the plastic hook (stored in the
holder at the side of the unit) to lift up a cassette
just far enough to be able to grab it and place it into
a tray.
3
Open each cassette carefully and remove the gels and
membranes. Label each membrane and indicate the
sample side. Lift membrane(s) with blunt forceps and
air dry, or follow the instructions accompanying your
protocol.
4
Discard the blotting paper, but reuse the sponges.
5
Rinse the unit immediately after use. (See the Care and
maintenance section on the next page.)
Note: It is a good idea to stain the
gel to determine the completeness
of the transfer.
Note: Do not store used buffer
with transfer tank. Chill buffer to
10 °C before reuse.

• p12
Care and maintenance
Cleaning
• Do not autoclave or heat any part above 45 °C.
• Do not expose to alcohols or organic solvents!*
• Never use abrasive detergents.
• If using radioactive reagents, decontaminate
the unit with a cleaning agent such as Contrad
70™ or Decon 90™.
Rinse the tank, cassettes, and sponges with
distilled water immediately after each use. Allow
the unit to air dry completely. Periodically wash
with a dilute solution of a mild detergent.
Removing the electrode panel(s)
For more thorough cleaning or to replace
damaged electrodes, remove each electrode panel
by unscrewing the retaining screw far enough to
allow the panel to slide out. Use the hook on the
side panel to pull the electrode panel up (do not
pull the panel up by the banana plug). Take care
to not stretch or break the platinum wire when
handling the panel.
* Use ≤20% methanol (methyl
alcohol) in transfer buffers is the
only exception.