Amersham Biosciences TE 70 User manual

user manual
um 28-4025-91/Rev. A0/09-05
ECL Semidry Blotters
TE 70 Semi-dry transfer unit
TE 77 Semi-dry transfer unit
TE 70 PWR Semi-dry transfer unit
TE 77 PWR Semi-dry transfer unit

Page finder
Important user information . . . . . . . . . . . . . . . . . . ii
WEEE information . . . . . . . . . . . . . . . . . . . . . . . . v
1. ECL Semi-dry blotters: Description. . . . . . . . . . . . . .1
Unpacking . . . . . . . . . . . . . . . . . . . . . . . . . . . 2
Specifications . . . . . . . . . . . . . . . . . . . . . . . . . 3
Important information . . . . . . . . . . . . . . . . . . . 4
2. Operating instructions. . . . . . . . . . . . . . . . . . . . . . .5
ECL Reagents . . . . . . . . . . . . . . . . . . . . . . . . 14
3. Care and maintenance . . . . . . . . . . . . . . . . . . . . .15
4. Troubleshooting . . . . . . . . . . . . . . . . . . . . . . . . . .16
5. Electrotransfer notes. . . . . . . . . . . . . . . . . . . . . . .19
6. Bibliography. . . . . . . . . . . . . . . . . . . . . . . . . . . . .21
7. Customer service information . . . . . . . . . . . . . . . .22
8. Ordering information . . . . . . . . . . . . . . . . . . . . . . .23
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Important user information
Please read this entire manual to fully understand the safe and effective
use of this product. Should you have any comments on this manual,
we will be pleased to receive them via email at
[email protected] or at:
Amersham Biosciences Corp.
800 Centennial Avenue
PO Box 1327
Piscataway, NJ 08855 USA
Amersham Biosciences reserves the right to make changes in the speci-
fications without prior notice.
Warranty and liability
Amersham Biosciences guarantees that the product delivered has been
thoroughly tested to ensure that it meets its published specifications.
The warranty included in the conditions of delivery is valid only if the
product has been installed and used according to the instructions
supplied by Amersham Biosciences.
Amersham Biosciences shall in no event be liable for incidental or
consequential 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.
Renseignements importants d’utilization
Pour une bonne compréhension et une utilisation en sécurité maximale,
il convient de lire entièrement ce manuel. Tous vos commentaires sur ce
manuel seront les bienvenus et veuillez les adresser à:
Amersham Biosciences Corp.
800 Centennial Avenue
PO Box 1327
Piscataway, NJ 08855 USA
[email protected]
Amersham Biosciences se réserve le droit d’effectuer des modifications
de ces spécifications sans aucun préavis.
Garantie et responsabilité
Amersham Biosciences 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 condi-
tions de livraison n’est valable que si le produit a été installé et utilisé
conformément aux instructions fournies par Amersham Biosciences.
La société Amersham Biosciences ne sera en aucun cas responsable de
tout dommage causé directement ou indirectement par toute utilisation
incorrecte ou non approuvée du produit ou découlant 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 limitation quant
à la nature de ces dommages.
um 28-4025-91 • pii

Información importante para el usuario
Para comprender el producto y utilizarlo con seguridad es necesario leer
este manual en su totalidad. Si desearan hacer algún comentario sobre
este manual, tengan la amabilidad de remitirlo a:
Amersham Biosciences Corp.
800 Centennial Avenue
PO Box 1327
Piscataway, NJ 08855 USA
[email protected]
Amersham Biosciences se reserva el derecho a modificar las especifica-
ciones sin previo aviso.
Garantía y responsabilidad
Amersham Biosciences garantiza que el producto entregado ha sido
probado a fondo para comprobar el cumplimiento de las especificacio-
nes publicadas. La garantía incluida en las condiciones de entrega sólo
es válida si el producto se ha instalado y utilizado de acuerdo con las
instrucciones entregadas por Amersham Biosciences.
Amersham Biosciences 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.
Wichtige benutzerinformationen
Für ein vollständiges Verständnis und eine sichere Handhabung dieses
Produktes ist es notwendig, daß der Benutzer dieses Handbuch voll-
ständig durchliest. Wenn Sie Anmerkungen zu diesem Handbuch haben,
dann senden Sie diese bitte an:
Amersham Biosciences Corp.
800 Centennial Avenue
PO Box 1327
Piscataway, NJ 08855 USA
[email protected]
Amersham Biosciences behält sich das Recht vor, die Spezifikationen
ohne vorhergehende Ankündigung zu ändern.
Gewährleistung and haftung
Amersham Biosciences garantiert, daß das gelieferte Produkt sorgfältig auf
die Einhaltung der veröffentlichten Spezifikationen getestet wurde. Die in
den Lieferbedingungen näher erläuterten Gewährleistungsansprüche gelten
nur dann, wenn das Produkt gemäß den von Amersham Biosciences gelief-
erten Anweisungen installiert und benutzt wurde.
Amersham Biosciences übernimmt keinerlei Haftung für Schäden oder
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eine fehlerhafte oder unsachgemäße Verwendung des Produkts verursa-
cht wurden.
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Informazioni importanti per l’operatore
Per un utilizzo sicuro del prodotto, leggere attentamente l’intero conte-
nuto del presente manuale. Si prega di inviare eventuali commenti al
presente manuale a:
Amersham Biosciences Corp.
800 Centennial Avenue
PO Box 1327
Piscataway, NJ 08855 USA
[email protected]
Amersham Biosciences si riserva il diritto di apportare modifiche ai dati
tecnici senza preavviso.
Garanzia e responsabilitá
Amersham Biosciences 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 Amersham Biosciences.
Amersham Biosciences 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.
um 28-4025-91 • piv

• pv
Waste Electrical and Electronic Equipment
(WEEE) information
This symbol indicates that the waste of electrical and elec-
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um 28-4025-91 • pvi

1. ECL™ Semi-dry blotters:
Description
The TE 70, TE 77, TE 70 PWR, and the TE 77
PWR semi-dry blotters rapidly transfer proteins
from polyacrylamide gels onto a membrane by
means of a low current and low voltage electro-
transfer with minimal Joule heating. Most trans-
fers are complete in one hour or less.
The smaller TE 70 unit transfer surface is
14 × 16 cm, suitable for transferring standard
gels, including those from the SE 600 Ruby and
the SE 400. The larger TE 77 transfer surface is
21 × 26 cm, suitable for transferring gels from
the Ettan DALT systems.
The TE 70 and TE 77 have safety circuits built
into the instrument that limit the voltage to 30 V
and the current to 0.5 A. The circuits protect the
user from unnecessary electrical hazards.
The TE 70 PWR and the TE 77 PWR 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 moni-
tor the transfer stack resistance, and can stop a
transfer if large changes indicate that the buffer
system is depleting.
Multiple gels can be transferred at the same time
by placing several small gels of the same thickness
side by side, or by stacking two gels vertically in
a carefully constructed multi-layered stack.
The electrodes are made of the best possible
materials to ensure the units last a long time.
• p1
Fig 1. Semi-dry transfer unit
main components.
power
cord
top electrode
power
transformer
lower electrode
TE 77 PWR
power
cord
top electrode
lower electrode
TE 70

Unpacking
Unwrap all packages carefully and compare
contents with the packing list, making sure all
items arrived. If any part is missing, contact
Amersham Biosciences. 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.
um 28-4025-91 • p2
The TE 70 and TE 77
require a power supply
with a minimum rating of
0 – 500 mA, 0 – 30 V.

Specifications
TE 70 TE 77 TE 70 PWR TE 77 PWR
Electrode size (cm) 14 × 16 21 × 26 14 × 16 21 × 26
Output:
Voltage 30 V 30 V 30 V 30 V
Current O.5 A O.5 A O.5 A O.5 A
Inputs:
Voltage 100 – 240 V 100 – 240 V
Current 0.7 A 0.7 A
Minimum transfer stack thickness:
TE 70, TE 70 PWR 3.2 mm
TE 77, TE 77 PWR 2.8 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
• p3
This declaration of conformity
is only valid for the instrument
when it is:
• used in laboratory locations.
• used as delivered from
Amersham Biosciences except
for alterations described in
the user manual.
• connected to other CE labeled
instruments or products
recommended or approved by
Amersham Biosciences.

Important information
• The electric components in the transfer unit base
must not become wet. Do not immerse the unit in
water. Rinse only the electrodes with distilled water
before and after use. (Refer to the Care and mainte-
nance section for cleaning instructions.)
• Be sure to use enough buffer-soaked sheets of blot-
ting paper on both sides of the membrane/gel stack
so that the buffer does not become depleted during
the transfer.
• 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
Amersham Biosciences may be used for operating,
maintaining, and servicing this product.
Informations importantes
• Les composants électriques dans la sûreté
enclenchent le logement de la base d’unité de
transfert ne doit pas devenir mouillé. Ne pas
immerger de partie de l’unité dans l’eau. Rincer
seulement les électrodes avec l’eau distillée avant
l’usage. (Se référer à la section de Soin et entretien
pour nettoyer d’instructions.)
• Etre sûr d’appliquer assez de tampon feuilles
trempées de tacher de papier sous la membrane et
par-dessus le gel pour empêcher la pile du séchage
hors. Les dommages irréparables à l’unité résult-
eront si la pile sèche hors et l’unité est permise de
surchauffer.
• Si cet équipement est utilisé dans une manière pas
spécifié par le fabricant, la protection fournie par
l’équipement peut être altérée.
• Seulement les accessoires et les parties ont
approuvé ou fourni par Amersham Biosciences sont
recommandés pour l’utilisation, l’entretien et répa-
ration de cet appareil.
um 28-4025-91 • p4
English
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Français
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2. Operating instructions
To transfer proteins, prepare the unit, assemble
the stack, and connect to a power supply if
necessary. Then run the transfer for the required
amount of time. Each step is described below.
Assemble the transfer stack
1
Prepare the unit by rinsing the anode and cathode
with distilled water.
2
Prepare the gel
Cut away the wells and/or stacking gel section.
Equilibrate the gel in transfer buffer if required.
3
Prepare the transfer stack
Cut the blot paper and transfer membrane to the
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 the masks.
Place the mask in the base of the unit, centering
the opening.
• p5
Note: Take care to place the gel
correctly on the first try because
proteins begin to transfer imme-
diately; once transfer has begun,
moving the gel will distort results
or cause “shadow” bands on
the blot.

4
Prepare the blotting paper
For each gel, cut at least 6 pieces of blotting paper
the same size as the gel or slightly smaller.
Gauge the thickness or number of blotting paper
layers according to the amount of buffer required; up
to 300 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.
Saturate at least 3 pieces of the blotting paper with
transfer buffer. One by one, center each sheet on the
lower electrode and remove all trapped air by rolling a
clean pipet or roller from the center toward the edges.
5
Prepare the membrane
Hybond membranes are recommended. (See related
products section).
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
types in transfer buffer for 2– 5 minutes.
um 28-4025-91 • p6
Note: Each 21 × 26 cm blot paper
will absorb approximately 50 ml
of transfer buffer. Each 14 × 16
cm blot paper will absorb 20 ml
of buffer.
Note: Always wear gloves when
handling membranes to avoid
leaving fingerprints.

6
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.
c) Cover the gel with three layers of saturated
blotting paper.
• p7
Important! Stack each layer with
care, with edges parallel. As each
layer is added, remove all air pock-
ets 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.
Mask (opening approx. 2 mm smaller
than the gel on all sides). Optional.
Blotting paper
(2–3 sheets)
Blotting paper
(2–3 sheets)
Gel
Membrane
These stack compo-
nents are the same
size as the gel or
slightly smaller
Fig 2. Transfer stack for a
single gel.

um 28-4025-91 • p8
Multiple gels: Either lay gels of the same thickness
side-by-side (Fig 3), or stack 2 sandwiches layered as
shown (Fig 4).
For best results, the transfer stack should be centered
in the electrode panels.
If two gels are stacked, separate them with porous
cellophane — not plastic wrap! (Cellophane permits
electric current to pass but stops proteins.) Cut the
cellophane slightly smaller than the gel and wet with
transfer buffer. Several sheets of buffer-soaked blot-
ting paper on each gel provides electrical continuity.
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, most
importantly, the characteristics of
the protein. The gel closest to the
anode generally transfers the most
completely. It is preferable to lay
gels side-by-side rather than
stacking them.
Fig 3. Transfer stacks for gels
placed side-by-side.
Note: There must be no electrical
contact between the two stacks.

• p9
Blotting 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.

um 28-4025-91 • p10
Electrotransfer
TE 70 and TE 77
1
The power supply should be switched off and both the
current and voltage controls set at zero
Then plug the color-coded leads from the base of the
transfer unit into the power supply jacks, matching
red to red and black to black. Do not reverse polarity.
(Red is positive [+] and black is negative [–].)
2
Set the power supply current
The maximum current setting should not exceed
0.8 mA/cm2 of the gel surface. If transferring several
layers of gels, a longer transfer time may be required.
Use the graph below to quickly find the current
setting for your gel size, or calculate the gel area
(cm2) and multiply it by 0.8 mA/cm2.
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 empirically.
cm2
Gel
Size
maximum mA
14 × 24
14 × 16
8 × 10
8 × 7
Fig 5. Recommended current
settings for different gel sizes.
Note: Generally, smaller
fragments transfer more
quickly than larger ones.
Note: Transfers exceeding one
hour will require additional
sheets of buffer saturated blot
paper in the transfer stack.

• p11
TE 70 PWR and TE 77 PWR
Turn the instrument on using the POWER
button on the front keypad.
The instrument works by setting the current and
time, and then starting the transfer. During the
transfer, the voltage can be displayed. The trans-
fer can not be set to run at constant voltage.
The DISPLAY MODE BUTTON toggles
between the current (mA), time (hour:minutes)
and the voltage (V).
The Up and DOWN arrows to change the value
for the current and time.
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 to
run continuously until manually
turned off by the user.
transfer
start/stop
up/down arrows
adjust value
display mode
cycles LED display
power
ON/OFF
Fig 6. TE 77 PWR keypad.

um 28-4025-91 • p12
During the transfer
TE 70 PWR and the TE 77 PWR
A red LED will appear in the lower right corner
of the Display when voltage is applied to the
transfer stack.
Both the Time and the Current can be changed
as the transfer progresses. Select mA or time
using the DISPLAY MODE button, and the
UP and DOWN to change the value. While the
value is being changed, the LED will blink. After
about 10 seconds the LED will stop flashing and
show the real time run parameters.
Buffer Depletion
One of the more common failure modes for a
Western transfer is buffer depletion. Buffer
depletion leads to changes in pH and overheating,
both of which are detrimental to the transfer.
The TE 70 PWR and the TE 77 PWR instru-
ments 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 prob-
lems, like burning of the transfer stack.
If this condition happens, the error message
“dry” will be on the display. The instrument has
stopped the transfer. If desired, the cover can be
removed, and buffer can be added to the transfer
stack. Replace cover, and press any key (except
Power ON/OFF) to continue.
For future transfers, add more layers of buffer
soaked blot paper.
Red LED
Dry error message.