miniVE - GE Healthcare Life Sciences

miniVE - GE Healthcare Life Sciences
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miniVE
Electrophoresis and Electrotransfer Unit
User Manual
80-6420-86
Rev. A/5-98
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F u n c t i o n
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D e s c r i p t i o n
Hoefer miniVE Electrophoresis Unit Function
and Description
The Hoefer miniVE vertical electrophoresis system performs vertical gel electrophoresis on mini-format gels. The basic unit includes two electrophoresis modules. Each module holds one gel sandwich, 10 cm wide and 8 to 10.5 cm long.
One gel can be cast in place on the electrophoresis module.
A wide range of accessories, ordered separately (see Section 5), lends the miniVE
a high degree of versatility. These include:
✛
a large selection of combs and spacers,
✛
adaptor sleeves to run Novex precast gels,
✛
and a blot module, which converts the miniVE into a mini blotting unit. (See
page 17 for instructions.)
Figure 1.1
Main components
Color-coded leads connect the
electrodes to the power supply.
Lid
Electrophoresis module
Banana plug connectors (2)
Power supply minimum
ratings:
50 mA, 250 V
Constant current or
constant voltage
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1.1 Specifications
Electrophoresis
Gel sandwich size
Max. tank volume
Max. voltage
Max. wattage
10.0 cm wide × 8 to 10.5 cm long
1.7 liters with one module in place
1.2 liters with two modules in place
600 V\
25 W per electrophoresis module
Electrotransfer
Max. volume (blot module)
350 ml per module
Max. tank volume (for passive cooling) 1.7 liters with one module in place
1.2 liters with two modules in place
Max. wattage
15 W per blot module
Max. current
400 mA
miniVE specifications
Max. operating temperature
Chemical compatibility
75 °C
For use only with dilute aqueous solutions
between pH 2 and pH 12. Not compatible
with organic solvents or concentrated alcohols, acids, bases, and oxidizing agents.
Environmental operating conditions
Indoor use: 4–40 °C
Humidity up to 80%
Altitude up to 2000 m
Installation category
II
Pollution degree
2
Dimensions (w × d × h)
Weight (tank, lid, and two gel modules)
19.2 × 17.2 × 18.8 cm (7.6 × 6.8 × 7.4 in.)
1.2 kg (2.65 lbs)
Product certifications
EN61010–1, UL3101–1, CSA C22.2 1010.1, CE
This declaration of conformity and the warranty are only valid when the instrument is:
◗ used in laboratory locations,
2
◗
used within the conditions specified in the User Manual,
◗
used as delivered from Amersham Biosciences except for alterations
described in the User Manual, and
◗
connected to other CE labeled instruments or products recommended or
approved by Amersham Biosciences.
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1.2 Important Information
1.2 Informations importantes
➧
The safety lid must be in place before
connecting the power leads to a power
supply.
➧
Le couvercle de sécurité doit être en place avant
de brancher les prises au générateur.
➧
Turn all power supply controls off and
disconnect the power leads before
removing the safety lid.
➧
Eteindre le générateur et débrancher les prises
avant d’enlever le couvercle de sécurité.
➧
Do not operate with the gel or buffer
temperature above 75 °C. Overheating
will cause irreparable damage to the
unit! Chilled buffer provides passive
cooling that can help control overheating
to some degree.
➧
Ne pas travailler avec un gel ou un tampon
dont la température dépasse 75 °C. Une surchauffe causerait des dommages irréparables à
l'unité d'électrophorèse ! En dessous de conditions normales, veuillez à prévenir d'une surchauffe en refroidissant le tampon avant l'utilisation.
➧
To clean the safety lid, wipe with a damp
towel or briefly rinse the underside only.
Do not immerse.
➧
Pour nettoyer le couvercle de sécurité, essuyer
avec un chiffon humide ou rincer uniquement
le dessous. Ne pas immerger.
➧
Do not autoclave or boil this unit or any
of its parts.
➧
Ne pas autoclaver ou stériliser cette unité, ni
aucune de ses pièces détachées.
➧
Do not remove the screws in the clamps;
four full turns in the counterclockwise
direction are sufficient to be able to open
the clamps.
➧
Ne pas enlever les visses des pinces ; pour
ouvrir les pinces, il suffit de tourner quatre fois
dans le sens inverse des aiguilles d'une montre.
➧
If this equipment is used in a manner not
specified by the manufacturer, the protection provided by the equipment may be
impaired.
➧
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.
➧
Only accessories and parts approved or
supplied by Amersham Biosciences
may be used for operating, maintaining, and servicing this product.
➧
Seulement les accessoires et piéces detachées
approuvés ou fournis par Amersham
Biosciences sont recommandés pour l’utilisation, l’entretien et réparation de cet
appareil.
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1.3 Unpacking
4
1.
Unwrap all packages carefully and compare contents with the packing list,
making sure all items arrived. If any part is missing, contact your local sales
office. 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 for repacking should it become necessary to return the unit.
2.
Prior to use, wash the tank and module with a dilute solution of non-abrasive laboratory detergent. Thoroughly rinse first with water and then with
distilled water.
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The Electrophoresis Module
This section describes the use of the electrophoresis module. For instructions
on using the blot module, see page 17.
2.1 Preparing the gel
Both self-cast and precast gels of the following dimensions fit the electrophoresis
module: 10 cm wide, 8 to 10.5 cm long, 0.75 to 1.5 mm thick. For instructions
for precast gels, see section 2.1.2.
2.1.1 Self-cast gels
One single gel can be cast on the module. To cast several gels, a multiple gel caster such as the Hoefer SE 215, SE 235 Mighty Small 4-Gel, SE 245 Mighty Small
Dual Gel Caster, or SE 275 Mighty Small Multiple Gel Caster is required (see
ordering information).
Clamp screws (4)
Figure 2.1
Module in the closed
position
Figure 2.2
Module in the open
position
Position the module to
accept the gel sandwich.
Each of the three sealing
elements is hinged, and
must be placed in the open
position:
a) Release the sealing plate
by applying gentle pressure
to both tabs as indicated
by the arrows. Then, holding the tabs, move the
plate into the fully open
position.
b
a
Sealing plate
c
b) Loosen all four screws
4–5 turns in the counterclockwise direction.
Note
The sealing plate has
three positions: closed
(sealed for casting),
half open (for electrophoresis), and fully
open (for placing the
gel sandwich).
H o e f e r
1.
c) Swing the clamps outward.
Lay the module flat on a
work surface.
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Clamps (2)
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2.
Figure 2.3
Gel sandwich assembly
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Assemble the gel sandwich.
Choose one notched plate, one rectangular glass
plate, and two spacers. Use only unchipped plates
to prevent leaking.
Assemble the gel sandwich as shown: the notch
is at the top of the sandwich and the spacer
ridges align along the glass plate edges on the
sides of the sandwich.
IMPORTANT: PROPER ALIGNMENT IS ESSENTIAL IN STEPS 3 AND 4 TO PREVENT
LEAKAGE. TAKE CAREFUL NOTE OF THE ALIGNMENT TIPS.
3.
Place the sandwich on the module.
Take care to “square” the three sealing sides of the sandwich. This can be
done by holding the sandwich like a deck of cards and gently tapping the
bottom against a flat surface.
Tip: Once the sandwich is carefully aligned, handle it with the flat sides
firmly between your thumb and fingers, near the notch.
Notched plate side down, lay the sandwich on the module, fitting it within
the guides at both sides (a) and against the guide feet at the bottom (b).
The notched plate is against the
module, with the notch at the top.
a
a
Figure 2.4
Placing the sandwich
b
b
The gel sandwich fits within guides at the
sides and aligns flush against the guide feet
at the bottom of the module.
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4.
Figure 2.5
Positioning the clamps
Figure 2.6
Checking the alignment
Figure 2.7
Misalignments will
cause leakage
Figure 2.8
Final assembly
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Seal the sandwich.
a
a) While gently holding the
sandwich against the module,
swing one clamp into position
over the spacer, taking care not
to bump the sandwich out of
alignment. Turn each screw
(alternating to keep the pressure even) until the clamps are
loosely secured and will allow
the spacers to be adjusted, if
necessary. Repeat on the other
side.
b) Important: Check the
alignment at the bottom of the
sandwich.
If the spacers and glass plates
are not perfectly aligned
against the stops, use the stiff
end of the Hoefer Wonder
Wedge provided to press
against the edges of the spacer
and glass plates to position
them flush against the guide
foot.
c) Complete clamping by tightening each screw firmly, hand
tight. Do not overtighten, as
the plates may crack. Check
the spacer alignment.
Tip: A test for alignment
involves passing a corner of the
Wonder Wedge across the bottom edge of the spacers and
glass plates. If an edge “catches,” realign. Check both sides.
b
Important
Check the bottom edges
for proper alignment
against the guide feet.
✗
✗
The spacer must
not protrude out of
the sandwich.
The glass plate should
not be resting on the
head of the spacer “T”.
Engage each tab in
the topmost notch.
c
d
d) Lock the sealing plate into
the closed position by engaging
each tab in its topmost notch.
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Figure 2.9
Convenient module
position for pouring the
gel
5.
If a counterweight is
required for balance,
either fill the tank or
hang the second module on the other side.
6.
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Hang the module from the side
of the tank or stand it on the
benchtop to cast the gel.
M o d u l e
Tabs on the narrow side of the
tank fit into slots in the module.
Prepare the monomer solution
and pour the resolving gel.
Caution: Acrylamide is a neurotoxin. Always wear gloves and
observe all laboratory safety procedures.
Pipet the solution into the sandwich slowly so that it flows
along a spacer, taking care not to
trap any air pockets.
Tip
Approximately 10 ml of
monomer solution is
required to cast one
1 mm thick gel,
10×10.5 cm plates.
No stacking gel: fill the solution to the desired level and then insert a comb
(at a slight angle) into the sandwich, taking care not to trap air under the
teeth.
For a 1 cm stacking gel below the wells, fill to 3 cm below the top of the
rectangular glass plate. Overlay each gel with a thin layer of water-saturated
n-butanol, water, or diluted gel buffer to prevent exposing the monomer
solution to oxygen. Apply the overlay solution (100 µl) slowly to one side of
the sandwich, near the spacer, using a glass syringe fitted with a 22-gauge
needle. Allow the solution to flow across the surface unaided.
7.
Allow a minimum of one hour for the gel to polymerize.
8.
If a comb is in place, remove it by carefully pulling on the comb while gently
rocking it back and forth to break the vacuum. Rinse the wells with electrophoresis buffer to remove any unpolymerized acrylamide.
If an overlay was applied, rinse the sandwich several times with double distilled water to remove it.
Invert the module to drain.
For instructions on pouring a stacking gel, see section 2.1.3.
9.
8
If the gel is ready for electrophoresis, move the sealing plate into the “half
open” position: Apply gentle pressure to both tabs and lock them into the
lower notch.
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2.1.2 Precast gels
1.
Novex gel cassettes only: Adaptor
sleeves, ordered separately, must be
installed on each clamp. Slide an adaptor sleeve on each pressure plate from
the bottom, as shown in Figure 2.10.
The sleeve fits in only one orientation,
and snaps into place when fitted properly.
Figure 2.10
Installing adaptor
sleeves
Figure 2.11
Securing the cassette
H o e f e r
See Section 2.1.1, step 1 to prepare the
electrophoresis module.
2.
Follow the manufacturer’s instructions
to prepare the gel for electrophoresis.
This may involve removing tape or
breaking off the sealing edge from the
bottom of the cassette.
3.
Remove the comb and rinse the wells
with electrophoresis buffer to remove
any unpolymerized acrylamide.
Tip: To aid in sample loading, mark
the well locations with a laboratory
marking pen.
Slide the adaptor sleeve
over the pressure plate.
a
a
4.
Position the cassette on the module.
Orient the cassette so that the notched
side is against the gasket, and the wells
are at the top of the module. Center
the cassette within the guides at both
sides of the module (a).
5.
Secure the cassette. Swing each clamp
into position over the sides of the cassette. Tighten each screw, alternating to apply even pressure until the cassette
is secure. The gasket around the upper buffer chamber should be fully compressed to provide a seal, but the screws should not be tightened to the point
that pressure stresses the cassette.
6.
Check that both gel surfaces will contact buffer. Novex gels: check that the
bottom gel-contact slot is exposed.
7.
Move the sealing plate into the “half open” position to prepare for electrophoresis: Apply gentle pressure to both tabs and lock them into the
lower notch.
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2.1.3 Cast the stacking gel
1.
If the gel has wells, skip to Section 2.2.
2.
The gel sandwich or cassette should already be in place on the module. To
ensure seamless contact between the resolving and stacking gels, remove
residual liquid by blotting one corner of the gel with a lint-free tissue.
3.
Prepare the stacking gel monomer solution.
Note
Stacking gel resolution
is optimal if the gel is
poured just before
electrophoresis.
10
Tip: To calculate the volume, measure the distance, in cm, from the top of
the resolving gel to the notch in the glass plate. (This should be at least 2
cm.) Multiply this distance by the gel width (8.3 cm) and the gel thickness
(cm) for the required volume (ml).
4.
Deaerate the stacking gel monomer solution, add catalyst and initiator and
then pour. Use a pipette to deliver the solution into one corner of the plate,
taking care not to trap any bubbles. Insert a comb (at a slight angle to prevent trapping air) into the sandwich, allowing the comb sides to rest on the
spacers.
5.
Allow a minimum of one hour for the gel to polymerize.
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2.2 Final assembly
1.
Figure 2.12
Preparing for electrophoresis
Tip: To aid in loading samples, either mark the well
location with a laboratory
marking pen or wet the welllocating decal and apply it to
the front of the glass plate so
that the appropriate edge outlines the sample wells.
Note for preparative
combs:
The side wells for
standards are the
same size as in the
10-well comb.
2.
Lower each module into the
tank, seating it in the locating
slots. The module seats properly in only one orientation:
with the banana plugs toward
the center of the tank, the gel
facing outward.
3.
Add the appropriate amount
of electrophoresis buffer to the
tank.
Note
Avoid wetting the
banana plugs with
electrophoresis buffer.
A protective film of
GelSeal™ (supplied)
can be applied to the
plugs as a corrosion
barrier.
Make sure the sealing plate is
in the “half open” position.
The arrow in Figure 2.12 indicates the correct position.
The half open
position for
electrophoresis
Upper buffer
chamber
General guidelines: add 1.2–1.6 liters of buffer to the tank when only one
module is in place, and 1.1–1.4 liters when two modules are in place.
The minimum and maximum levels are marked. The minimum level ensures
that the lower electrode, which is approximately 2 cm from the bottom of
the module, is completely submerged. Verify that it is. The maximum level
prevents buffer in the tank from entering the upper buffer chamber. Also
verify this.
4.
Add the appropriate amount of electrophoresis buffer to the upper buffer
chamber.
Fill the upper buffer chamber to a level 3–5 mm above the notched plate.
For a 10.5 cm long gel approximately 100 ml will be required, and for a
shorter gel, approximately 75 ml.
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5.
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Prepare and apply the sample.
Increase liquid sample density with 10% glycerol or sucrose. Add a tracking
dye such as phenol red or bromophenol blue.
Underlay the sample into the wells using a micro-pipet or fine-tipped
microsyringe. Table 1 shows the volume of sample required for different
numbers of wells and comb thicknesses.
Note: The amount of protein sample added to each well depends on both
the sensitivity of the staining method and the distribution of protein among
separate bands. With Coomassie Blue, it is possible to detect 1 µg in a single
band; with the more sensitive silver stains, it is possible to detect as little as
10 ng.
Table 2.1
Well capacities
Volume of sample (µl) per 1 mm depth
No. of wells
5
9
10
15
18
6.
Figure 2.13
Fully assembled
Comb thickness (mm)
0.75
1.0
1.5
9.5
3.6
2.2
12.7
5.8
4.8
2.9
2.9
19.1
7.2
4.4
Electrical connections.
Position the safety lid over the
unit and seat the lid so the banana
plugs engage the jacks in the lid.
The lid is symmetrical and fits in
either orientation.
Plug the color-coded leads into the
jacks of an approved power supply (red to red, black to black).
The minimum power supply rating
is 250 V, 50 mA, constant current
or constant voltage.
(Recommended power supply:
EPS 300.)
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2.3 Electrophoresis
For optimal resolution, electrophoresis should be started immediately after sample
loading.
Important
After initial monitoring,
do not leave the unit
unattended for more
than 30 minutes before
checking the buffer
level and the progress
of the bands.
Gels may be run at either constant current or constant voltage. For Laemmli SDS
separations, the recommended voltage range is 100–250 V and should not exceed
300 V. If running gels at constant current, the current should be 10–20 mA per
gel, depending on gel thickness (10 mA for 0.75 mm, 15 mA for 1.5 mm).
Check progress after 5 minutes, and again after half an hour, monitoring the position of the tracking dye. The run is complete when the tracking dye reaches the
bottom of the gel.
2.4 After electrophoresis
1.
Turn off the power supply and disconnect the leads.
2.
Remove the safety lid and lift out the module(s).
3.
Release each gel sandwich or cassette from the module: Move the sealing
plate to the fully open position by pressing inward on both tabs and guiding
the plate to open out. Then unscrew all four screws 4–5 turns in the counterclockwise direction. Swing the clamps outward.
4.
Remove the gel from the sandwich or cassette.
Important
Always disconnect the
high voltage leads
from the power supply
before removing the lid
from the unit.
Gently loosen and then slide away both spacers. Slip an extra spacer or the
Hoefer Wonder Wedge into the bottom edge (to prevent breaking the “ears”
of the notched plates) and separate the plates.
If using precast gels, follow gel manufacturer’s instructions.
Carefully lift the gel from the plate and lay it into a tray containing stain,
fixative, or transfer buffer.
5.
H o e f e r
Clean the unit as described in Section 2.5.
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2.5 Care and Maintenance
◗
Do not autoclave or heat any part above 75 °C.
◗
Do not immerse the safety lid in any liquid.
◗
Do not use organic solvents, strong or oxidizing cleaning solutions, abrasives, or strong acids or bases on any part of the instrument.
Immediately after each use, rinse the tank and modules with water and then rinse
thoroughly with distilled water. Handle the module with care to prevent damage
to the banana plugs. Allow to air dry.
Wipe the lid with a damp cloth. If necessary, briefly rinse the underside of the lid
with water.
Clean glass plates and spacers with a dilute solution of a laboratory detergent
such as RBS-35®, then rinse thoroughly with tap and distilled water. Glass plates
can be treated with (but not stored in) acid cleaning solutions. A final wipe with
isopropanol will remove any GelSeal residue.
2.6 Troubleshooting
Unusually slow (or fast) run
Reagent and solution factors
✓ Check recipes, gel concentrations, solutions, and dilutions.
(e.g., do not use Tris-HCl instead of Tris.)
✓
If the required pH of a solution is exceeded, do not back-titrate.
Prepare fresh buffer.
✓
Use only stock of the highest quality. Dispose of outdated acrylamide
solutions.
✓
Only use freshly deionized urea.
Voltage or current settings
✓ To increase or decrease the migration rate, adjust the voltage or
current by 25–50%.
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Stained sample collects:
Near the buffer front
✓ Protein or nucleic acid is not sufficiently restricted by the resolving
gel; increase the % T.
Near the top of the gel when the buffer front has reached the bottom
✓ The gel pore size is too small. Decrease the % T of the resolving
gel.
✓
Protein precipitates or the DNA becomes denatured. Decrease the
temperature at which the sample is prepared to 70 °C or less, and
limit exposure to heat to 1–2 minutes.
Smile effect on the buffer front
To reduce the running temperature:
✓ Prechill the buffer.
✓
Decrease the current or voltage setting. (Laemmli gel guidelines:
10 mA per 0.75 mm gel, 15 mA per 1.5 mm thick gel.)
✓
Conduct electrophoresis in the cold room.
✓
Fill the tank to the maximum (marked) buffer level.
Bands are skewed or distorted
Gel preparation
✓ Deaerate the stacking gel solution and avoid trapping air bubbles
under the comb teeth.
✓
Overlay the monomer solution with water-saturated n-butanol to
avoid forming an uneven gel surface.
Sample preparation
✓ Dialyze or desalt the sample.
✓
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Centrifuge or filter the sample to remove particulates.
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Poor band resolution
✓
Use only the highest quality reagents.
✓
Only use freshly deionized urea.
✓
Use only gels that were recently prepared.
✓
Check pH values of the resolving and stacking gel solutions. Do
not back-titrate buffers.
✓
Conduct the separation at a lower current or voltage setting.
Sample preparation
✓ Dialyze or desalt the sample.
✓
Reduce the sample volume or concentration.
✓
Improve dissociation of protein subunits by heating sample in SDS
sample buffer 1–2 minutes at 100 °C. Store on ice after heating.
✓
Store sample on ice before it is denatured.
✓
Add protease inhibitors if necessary to prevent proteolytic degradation
of sample.
✓
Add more mercaptoethanol or dithiothreitol; check sample treatment.
✓
Store samples to be frozen in aliquots to prevent repeated thawing. Store at -40 °C to -80 °C.
Protein streaks vertically
✓
Centrifuge or filter the sample to remove particulates.
✓
Dialyze or desalt the sample.
Bromophenol blue doesn’t sharpen into a concentrated zone in the stacking gel
16
✓
Pour a taller stacking gel. (For best results, allow a stacking gel
height of 2.5 times the height of the sample in the well.)
✓
Dispose of outdated acrylamide solutions and use only the highest
grade of acrylamide.
✓
When preparing samples, avoid using solutions with a high sodium
or potassium concentration.
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The Blot Module (ordered separately)
The Hoefer miniVE blot module, ordered separately, performs electrotransfers on
mini-format gels. Each module holds up to two gels, 8.2 cm wide and up to
10.4 cm long. One or two modules can be run at one time.
3.1 Assembly
1.
Prior to use, wash the tank and blot module with a dilute solution of nonabrasive laboratory detergent. Thoroughly rinse with water and distilled
water.
2.
Install the gaskets. Separate out two of the four strands of gaskets included
with each module.
Open the module by unlatching both tabs. Lay a gasket along the entire
groove around each cup half. Avoid stretching or twisting the gasket; the
length should just fit. Gently press into place.
Figure 3.1
Install a gasket into
each cup half.
Open the cup by releasing both tabs.
Once gaskets are in place, the cup will
pop open when the tabs are released.
A gasket fits into the groove around
three sides of each cup half.
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3.2 Preparation
3.2.1 Optional: Passive cooling
Chill approximately 2 liters of deionized water to 4 °C. (Filling the tank with
chilled water serves as a heat sink during electrotransfer.)
3.2.2 Prepare transfer buffer
Transfer buffer is required for stack assembly (≈250 ml) and to fill each module
(300–350 ml per module). The recipe for the commonly used Towbin buffer is
listed below. The Bibliography lists sources for other buffers.
Towbin buffer
(25 mM Tris, 192 mM glycine, 0-20% v/v methanol, pH 8.3, 1 liter)
Tris (FW 121.1)
Glycine (FW 75.07)
SDSa (FW 288.4)
25 mM
192 mM
0.1% (3.5 mM)
3.0 g
14.4 g
1.0 g
a
Optional: Adding SDS can improve transfer efficiency.
1.
Dissolve in 750 ml distilled water.
2.
Add methanol as required. Depending on the membrane type selected,
adding methanol can improve the transfer results. Because buffers containing
methanol may deteriorate if stored for long periods, add methanol just prior
to transfer.
3.
Bring to 1 liter with distilled water. Do not adjust the pH, which should be
between 8.2 and 8.4.
4.
Optional: Chill before use.
3.3 Prepare the transfer stack
Transfer the sample as soon as possible after electrophoresis to minimize sample
diffusion within the gel. Electrophoretic transfer can be performed on as many as
four mini gels at one time if two gels are placed in each of two modules.
1.
18
The transfer stack consists of the gel and membrane, filter paper, and three
packing sponges. The gel determines the size of the membrane and filter
paper: for each gel cut the membrane and two pieces of filter paper the
same size as the gel, but no larger than 8.5 × 10.5 cm.
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2.
Equilibrate the gel in transfer buffer for 10 minutes. Equilibration allows the
gel to swell or shrink before it contacts the transfer membrane and removes
excess buffer salts and detergents from the gel. Longer equilibration may
result in diffuse bands.
3.
The transfer membrane is prepared in two steps:
a. Pre-wet nitrocellulose or nylon membranes in distilled water, taking care
not to trap air bubbles: dip one end of the membrane into the buffer and
slowly submerge, allowing it to wet by capillary action. Pre-wet PVDF or
other hydrophobic membranes in methanol.
b. After prewetting, soak all membrane types in transfer buffer for 2–5 minutes.
4.
Wet the two pieces of filter paper in transfer buffer.
5.
Assemble the transfer stack so that molecules will migrate to the membrane.
For negatively charged macromolecules (such as proteins run in an SDS gel
and nucleic acids), assemble the transfer stack on the black cathode side.
Note: For best results, care should be taken to avoid trapping air bubbles as
each layer is applied. This is best accomplished by always establishing full
contact along one side and maintaining it as the layer is lowered into position.
See step 5 for proper orientation.
a. Center a packing sponge on the black cathode side.
b. Lay one piece of wet filter paper on the sponge.
c. Position the equilibrated gel on the filter paper. Wet
the gel surface with a few drops of transfer buffer.
d. Lay the membrane on the gel. Do not reposition the
membrane once it contacts the gel.
e. Lay one piece of wet filter paper on the membrane.
f. Lay two packing sponges on the filter paper.
A second transfer stack, if added, is placed
between these two sponges: Repeat steps b–e.
Figure 3.2
Transfer stack
assembly
The module is color
coded:
black = cathode (–)
red = anode (+)
f.
Important
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.
H o e f e r
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b.
c.
d.
e.
a.
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Check the position of the transfer stack. It should be centered on the electrode plate; no layer should be pinched when the module is closed. Fold the
empty half of the cup over the stack and press the halves together to snap
the module closed.
3.4 Final assembly
Note
Avoid wetting the
banana plugs with
electrophoresis buffer.
A protective film of
GelSeal™ (supplied)
can be applied to the
plugs as a corrosion
barrier.
1.
Pour 300–350 ml of transfer buffer into the top of the module. Tap the blotting cup lightly to dislodge any air bubbles in the packing sponges.
2.
Position the module(s) in the tank with the banana plugs toward the center
(the red side facing outward).
3.
Add deionized water to the tank: 1.7 liters for one module and 1.2 liters for
two modules.
Buffer temperature should not exceed 75 °C to avoid rapid evaporation.
Passive cooling is recommended if the transfer will be longer than one hour,
if biological activity must be protected, or if transferring nucleic acids. Chill
deionized water to (4 °C) before adding to the tank.
4.
Place the safety lid on the tank. (Either orientation fits and is correct.) Plug
the color-coded leads into the jacks of an approved power supply (such as
the EPS 300): red to red, black to black.
Figure 3.3
Final assembly
Color-coded leads connect
the electrodes to the power
supply.
Banana plug connectors
(2 per module)
20
The black cathode side (+)
faces toward the center.
The red anode side (−) faces
toward the outside tank wall.
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3.5 Electrotransfer
Note
Never leave the unit
unattended for more
than one hour once
electrotransfer has
begun.
Note
300–350 ml Towbin
buffer contains sufficient buffering capacity
for a transfer period of
up to 2 hours at
300 mA.
Electrophoretic transfer conditions for blotting proteins in Towbin buffer: 25 V
for 1–2 hours, 300–400 mA.
Important: We recommend programming the power supply to hold the current
setting constant to avoid possible overheating, especially if no passive cooling is in
place. (Buffer conductivity increases with increasing temperature, providing a positive feedback that results in rapid heating.) If the only programming option is to
hold the voltage setting constant, monitor and adjust the voltage to maintain the
current at or below 400 mA.
3.6 After electrotransfer
1.
Turn off the power supply and disconnect the leads.
2.
Remove the safety lid. Lift out the module(s) and drain by inverting over a
sink. Avoid wetting the banana plugs with buffer.
3.
Open the module. Remove the gels and membranes. Save the packing
sponges. Discard the blotting paper.
4.
Label each membrane and indicate the sample side. Lift the membrane(s)
with blunt forceps and allow to air dry.
5.
Rinse the unit immediately after use.
3.7 Care and Maintenance
◗
Do not autoclave or heat any part above 75 °C.
◗
Do not use organic solvents, strong or oxidizing cleaning solutions, abrasives, or strong acids or bases on any part of the instrument.
Immediately after each use, rinse the unit with water and then rinse thoroughly
with distilled water. Handle the module with care to prevent damage to the electrode plugs. Allow to air dry.
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3.8 Troubleshooting
Incomplete transfer
Blank areas on the membrane
✓
Remove all trapped air bubbles in the transfer stack; take especially great care during stack assembly to prevent air bubbles from
forming as each layer is placed.
✓
Check electrode continuity.
✓
Use a lower ionic strength buffer.
Molecules do not migrate out of gel
✓
Increase the field strength.
✓
Increase the transfer period. (Try doubling it.)
✓
Do not expose the gel to staining or fixing agents before transfer.
✓
Use a thinner gel.
✓
Reduce the gel acrylamide concentration.
✓
For proteins, use 3.5 mM SDS (0.1%) in the transfer buffer.
✓
Avoid using methanol in the protein transfer buffer or reduce the
amount to a minimum.
✓
Use reagent grade chemicals.
✓
Increase the length of time Southern blots are depurinated.
✓
Check the buffer pH. Most buffers should not be titrated; make
fresh buffer.
✓
Increase the net charge on the protein by changing to a transfer
buffer with a different pH. Lower pH (<6-7) increases the positive
charge on proteins; higher pH (>6-7) increases the negative
charge on proteins.
Diffuse band patterns
22
✓
Conduct the electrotransfer immediately after electrophoretic separation.
✓
Shorten or eliminate the equilibration step before electrotransfer,
or conduct equilibration in the cold room.
If the transfer buffer contains methanol (≥10%), however, equilibrate the gel for 30 minutes to allow it to shrink fully. Note: Gel
shrinkage may slow the migration of large molecules.
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✓
Take care that the gel does not shift once it contacts the membrane.
✓
Check that the preferred binding surface of the membrane faces
the gel.
Inefficient binding to membrane
Chemical parameters
✓
Fix or crosslink the molecule onto the membrane according to the
requirements of the nucleic acid, protein, or membrane type.
✓
Prepare protein transfer buffer without SDS.
✓
Verify the optimal amount of methanol required for the membrane
type and check the buffer solution. Add 10–20% methanol to the
transfer buffer to enhance binding to nitrocellulose.
Membrane parameters
H o e f e r
m i n i V E
✓
Wear gloves when handling membranes.
✓
Store membranes properly (e.g. protect from temperature
extremes and direct sunlight).
✓
If proteins pass through the selected membrane, try a different
type or one with a smaller pore size (0.10–0.20 µm).
✓
Place a membrane on both sides of the gel if different proteins
may be migrating in opposite directions.
✓
If the sample load may be exceeding the capacity of the binding
surface area, apply two membranes. If "blow through" occurs,
reduce the sample load.
23
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Bibliography
4.1 Polyacrylamide gel electrophoresis
Adams, L.D. and Gallagher, S.R., Two-Dimensional Gel Electrophoresis Using the O'Farrell
System. Current Protocols in Molecular Biology, 10.4.1–10.4.13 (1992).
Gallagher, S.R., and Smith, J.A., Electrophoretic separation of proteins. Current Protocols in
Molecular Biology. (F.A. Ausubel, R. Brent, R.E. Kingston, D.D. Moore, J.G. Seidman, J.A.
Smith, and K. Struhl, eds.) 10.2.1–10.2.21 (1991).
Laemmli, U.K., Cleavage of structural proteins during the assembly of the head of bacteriophage
T4. Nature. 227, 680–685 (1970).
Matsudaira, P.T. and Burgess, D.R., SDS microslab linear gradient polyacrylamide gel electrophoresis. Anal. Biochem. 87, 386–396 (1978).
Reisfeld, R.A., et al. Acidic buffer system for resolution of cationic proteins. Nature. 195, 281
(1962).
Sasse, J., and Gallagher, S.R., Staining proteins in gels. Current Protocols in Molecular Biology.
(F.A. Ausubel, R. Brent, R.E. Kingston, D.D. Moore, J.G. Seidman, J.A. Smith, and K.
Struhl, eds.) 10.6.1–10.6.8 (1991).
Weber, K., and Osborn, M., The reliability of molecular weight determinators by dodecyl sulfatepolyacrylamide gel electrophoresis. J. Biol. Chem. 224, 4406–4412 (1969).
4.2 Blotting
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).
Gershoni, J.M., and G.E. Palade Protein Blotting: Principles and Applications. Anal. Biochem.
131, 1–15 (1983).
Harlow, E. and Lane, D., Antibodies: A Laboratory Manual, Cold Spring Harbor Press, (1988).
Sambrook, J., et al. Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory
Press, B.23 (1989).
Towbin, H., Staehelin,T., and Gordon, J., Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc. Natl. Acad. Sci.
USA. 76, 4350–4354 (1979).
24
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C u s t o m e r
S e r v i c e
I n f o r m a t i o n
Customer Service Information
Technical Service and Repair
Amersham Biosciences offers complete technical support for all our products. If you have any questions about how to use this product, or would like to
arrange to repair it, please call or fax your local Amersham Biosciences
sales office or representative.
Important: Request a copy of the “Health and Safety Declaration” Form before
returning the item. No items can be accepted for servicing or return unless this
form is properly completed.
Ordering Information
Qty.
Code No.
Hoefer miniVE Basic: includes two gel modules, lid and tank.
1
80-6418-58
Hoefer miniVE, complete: includes 3 rectangular glass plates,
3 notched plates, 2 gel modules, lid, tank, 2 each 1.0 mm thick
10 well combs and 1.0 mm thick spacer sets.
1
80-6418-77
Adaptors for use with Novex precast gels
4/pk
80-6421-24
1
80-6418-96
Glass plates, 10 x 10.5 cm
5/pk
80-6150-87
Notched glass plates, 10 x 10.5 cm
5/pk
80-6150-49
Spacers, 0.75 mm thick
pair
80-6149-92
Spacers, 1.0 mm thick
pair
80-6150-11
Spacers, 1.5 mm thick
pair
80-6150-30
Comb; 5 well, 0.75 mm thick
1
80-6140-23
Comb; 5 well, 1.0 mm thick
1
80-6140-42
Comb; 5 well, 1.5 mm thick
1
80-6140-61
Comb; 9 well, 1.0 mm thick (microtiter)
1
80-6140-80
Comb; 10 well, 0.75 mm thick
1
80-6138-71
Comb; 10 well, 1.0 mm thick
1
80-6138-90
Comb; 10 well, 1.5 mm thick
1
80-6139-09
Comb; 15 well, 0.75 mm thick
1
80-6139-47
Blot Module, includes 3 Dacron sponges (¼" thick),
25 sheets of blotter paper
Accessories
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I n f o r m a t i o n
Comb; 15 well, 1.0 mm thick
1
80-6139-66
Comb; 15 well, 1.5 mm thick
1
80-6139-85
Comb; 18 well, 1.0 mm thick (microtiter)
1
80-6140-04
Comb; prep/ref., 0.75 mm thick
1
80-6141-56
Comb; prep/ref., 1.0 mm thick
1
80-6141-75
Comb; prep/ref., 1.5 mm thick
1
80-6141-94
80-6146-50
Gel Casters, Hoefer SE 200 Series
SE 245 Mighty Small Dual Gel Caster, 1 or 2 gels, 10 x 8, -10.5 cm
1
SE 215 Mighty Small Multiple Gel Caster, 5 to 10 gels, 10 x 8 cm
1
80-6142-51
SE 275 Mighty 4-Gel Caster, 2 to 4 gels, 10 x 8 cm
1
80-6151-06
SE 235 Mighty 4-Gel Caster, 2 to 4 gels, 10 x 10. 5 cm
1
80-6146-12
Electrophoresis Power Supplies
EPS 300 Power Supply, 300 V, 400 mA, 80 W
1
18-1123-97
Hoefer EPS 2A200 Power Supply, 200 V, 2000 mA, 200 W
1
80-6406-99
Gel Drying System
Hoefer EasyBreeze Air Gel Dryer 115 V
1
80-6121-61
Hoefer EasyBreeze Air Gel Dryer 230 V
1
80-6121-80
LMW Marker Kit
1
17-0446-01
HMW-SDS Marker Kit
1
17-0615-01
HMW Marker Kit
1
17-0445-01
Protein Molecular Weight Markers
PlusOne™ Chemicals
Acrylamide PAGE
250 g
17-1302-01
Acrylamide PAGE
1 kg
17-1302-02
1000 ml
17-1303-01
250 g
17-1300-01
1000 ml
17-1301-01
100 g
17-1304-02
1000 ml
17-1305-01
25 g
17-1311-01
TEMED
25 ml
17-1312-01
Acrylamide PAGE 40% solution
Acrylamide IEF
Acrylamide IEF 40% solution
N,N’ -methylenebisacrylamide
N,N’ -methylenebisacrylamide 2% solution
Ammonium persulfate
26
Tris
500 g
17-1321-01
Boric Acid
500 g
17-1322-01
EDTA, disodium salt
100 g
17-1324-01
Urea
500 g
17-1319-01
Silver Staining Kit, Protein
1
17-1150-01
Silver Staining Kit, DNA
1
17-6000-30
H o e f e r
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S e r v i c e
I n f o r m a t i o n
Amersham Life Science Products
Rainbow colored molecular weight markers (low molecular weight)
1
RPN755
Rainbow colored molecular weight markers (high molecular weight)
1
RPN756
Rainbow colored molecular weight markers (full range)
1
RPN800
ECL Protein molecular weight markers with Strep-HRP
1
RPN2280
ECL Western blotting molecular weight markers
1
RPN2107
ECL Plus Western blotting detection system
1
RPN2132
ECL Western blotting reagents (sufficient for 1000 cm2 of membrane) 1
RPN2109
ECL Western blotting reagents (sufficient for 2000 cm2 of membrane) 1
RPN2209
ECL Western blotting reagents (sufficient for 4000 cm2 of membrane) 1
RPN2106
ECL Western blotting system (kit; sufficient for 1000 cm2 of membrane)1
RPN2108
Hybond-C Super supported pure nitrocellulose, 12 x 10 cm
20/pk
Hybond-C nitrocellulose membrane, 12 x 10 cm
20/pk
RPN1210C
Hybond-C pure nitrocellulose membrane, 20 x 20 cm
10/pk
RPN2020W
Hybond-C pure nitrocellulose membrane, 20 cm x 3 m
roll
RPN203W
Hybond-C pure nitrocellulose membrane, 30 cm x 3 m
roll
RPN303W
10/pk
RPN2020F
Hybond-P, PVDF membrane, 20 x 20 cm
Hybond-P, PVDF membrane, 30 cm x 3 m
RPN1210G
roll
RPN303F
Hybond ECL nitrocellulose membrane, 20 x 20 cm
10/pk
RPN2020D
Hybond ECL nitrocellulose membrane, 6 x 8 cm
50/pk
RPN68D
roll
RPN303D
Hybond ECL nitrocellulose membrane, 30 cm x 3 m
Hyperfilm ECL, 12.5 x 17.5 cm
25/pk
RPN1674H
Blotter paper, 7 x 8 cm
25/pk
80-6211-48
Blotter paper, 9 x 10.5 cm
50/pk
80-6205-40
Nitrocellulose, 0.45 µm pore size, 9 x 10.5 cm
10/pk
80-6221-17
Nitrocellulose, 0.45 µm pore size, 33 cm x 3 m
roll
80-6221-55
Nitrocellulose, 0.20 µm pore size, 33 cm x 3 m
roll
80-6220-22
Nylon 66 Standard, 0.45 µm pore size, 33 cm x 3 m
roll
80-6221-93
Nylon Standard (GeneBind), 0.45 µm pore size, 20 cm x 3 m
roll
80-1247-87
Nylon 66 Plus (charged), 0.45 µm pore size, 33 cm x 3 m
roll
80-6221-74
1
80-6414-02
6/pk
80-6416-11
12/pk
17-6001-10
2-D Instruments and Accessories
IPGphor System 115/230 V (Order Strip Holders separately)
7 cm Strip Holder, complete for use
with Immobiline Drystrip IPG gels
Immobiline DryStrip, pH 4-7 L, 7 cm
H o e f e r
Immobiline DryStrip, pH 3-10 L, 7 cm
12/pk
17-6001-11
Immobiline DryStrip, pH 3-10 NL, 7 cm
12/pk
17-6001-12
m i n i V E
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ELECTROPHORESIS UNIT FUNCTION AND DESCRIPTION
1.1 SPECIFICATIONS . . .
1.2 IMPORTANT INFORMATION
1.3 UNPACKING . . . . .
1
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2. THE ELECTROPHORESIS MODULE
2.1 PREPARING THE GEL . . .
2.2 FINAL ASSEMBLY . . . .
2.3 ELECTROPHORESIS . . .
2.4 AFTER ELECTROPHORESIS
2.5 CARE AND MAINTENANCE.
2.6 TROUBLESHOOTING . . .
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3. THE BLOT MODULE
3.1 ASSEMBLY . . . . . . . .
3.2 PREPARATION. . . . . . .
3.3 PREPARE THE TRANSFER STACK .
3.4 FINAL ASSEMBLY . . . . . .
3.5 ELECTROTRANSFER . . . . .
3.6 AFTER ELECTROTRANSFER . .
3.7 CARE AND MAINTENANCE. . .
3.8 TROUBLESHOOTING . . . . .
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4. BIBLIOGRAPHY
24
5. CUSTOMER SERVICE INFORMATION
25
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Renseignements importants d’utilization
Français
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:
Deutsch
Amersham Biosciences
Marketing Department
654 Minnesota Street
San Francisco, CA 94107 USA
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 conditions 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.
Copyright© 1998 Amersham
Biosciences
Amersham Biosciences se réserve le droit d’effectuer des modifications de ces spécifications sans
aucun préavis.
Tous droits réservés. La reproduction, le stockage
dans un système de récupération d’informations ou la
transmission sous quelque forme que ce soit et par
quelque moyen que ce soit de la présente publication
en totalité ou en partie sont strictement interdits sans
autorisation préalable écrite de la société.
Wichtige Benutzerinformationen
Gewährleistung and Haftung
Für ein vollständiges Verständnis und eine sichere
Handhabung dieses Produktes ist es notwendig, daß
der Benutzer dieses Handbuch vollständig durchliest.
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
gelieferten Anweisungen installiert und benutzt wurde.
Ein Ausrufezeichen in einem gleichseitigen Dreieck soll den Benutzer auf die
Anwesenheit wichtiger Betriebs- und
Wartungsanweisungen in der dem Gerät
beiliegenden Dokumentation 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:
Amersham Biosciences
Marketing Department
654 Minnesota Street
San Francisco, CA 94107 USA
Amersham Biosciences behält sich das Recht
vor, die Spezifikationen ohne vorhergehende
Ankündigung zu ändern.
ii
Garantie et responsabilité
Amersham Biosciences übernimmt keinerlei
Haftung für Schäden oder Folgeschäden,
einschließlich, aber nicht begrenzt auf
Gewinneinbußen, Einkommensverluste, entgangene
Geschäftsabschlüsse, Verlust der Gebrauchsfähig-keit
oder andere Verluste, die wie auch immer durch eine
fehlerhafte oder unsachgemäße Verwendung des
Produkts verursacht wurden.
Copyright© 1998 Amersham Biosciences
Alle Rechte vorbehalten. Die vorliegende
Veröffentlichung darf nur mit vorhergehender
schriftlicher Genehmigung durch das Unternehmen
vervielfältigt, in einem Abrufsystem gespeichert oder
in irgendeiner Form oder mit irgendwelchen Mitteln
übertragen werden.
H o e f e r
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Informazioni importanti per l’operatore
Garanzia e responsabilitá
Per un utilizzo sicuro del prodotto, leggere attentamente l’intero contenuto del presente manuale.
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 .
Italiano
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:
Amersham Biosciences
Marketing Department
654 Minnesota Street
San Francisco, CA 94107 USA
Amersham Biosciences si riserva il diritto di
apportare modifiche ai dati tecnici senza preavviso.
Copyright© 1998 Amersham Biosciences
Tutti i diritti riservati. Nessuna parte della presente
pubblicazione può essere riprodotta, conservata in sistemi di gestione dati o trasmessa in alcun forma né
per nessuno scopo senza autorizzazione scritta del
produttore.
Información importante para el usuario
Garantía y responsabilidad
Para comprender el producto y utilizarlo con seguridad es necesario leer este manual en su totalidad.
Amersham Biosciences garantiza que el producto entregado ha sido probado a fondo para comprobar el cumplimiento de las especificaciones 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 .
Español
El signo de admiración en un triángulo
equilátero en el manual, advierte al
usuario sobre la presencia de instrucciones 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:
Amersham Biosciences
Marketing Department
654 Minnesota Street
San Francisco, CA 94107 USA
Amersham Biosciences se reserva el derecho a
modificar las especificaciones sin previo aviso.
H o e f e r
Amersham Biosciences non potrà essere ritenuta responsabile di incidenti 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.
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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.
Copyright© 1998 Amersham Biosciences
Reservados todos los derechos. No está permitida la
reproducción, ni el almacenaje en un sistema de recuperación, ni la transmisión de parte alguna de esta
publicación sin la autorización por escrito de la
empresa.
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Important user information
Warranty and Liability
Please read this entire manual to fully understand the
safe and effective use of this product.
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.
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:
Amersham Biosciences
Marketing Department
654 Minnesota Street
San Francisco, CA 94107 USA
Amersham Biosciences reserves the right to
make changes in the specifications without prior
notice.
iv
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.
Copyright© 1998 Amersham
Biosciences
All rights reserved. No part of this publication may be
reproduced, stored in a retrieval system or transmitted
in any form by any means, without permission in
written form from the company.
H o e f e r
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