Mini-PROTEAN®3 Dodeca
™
Cell
Instruction Manual
Catalog Number
165-4100
For Technical Service Call Your Local Bio-Rad Office or in the U.S. Call 1-800-4BIORAD (1-800-424-6723)
Table of Contents
Page
Section 1 General Information ................................................................................1
1.1 Introduction............................................................................................................1
1.2 Safety .....................................................................................................................1
1.3 Components ...........................................................................................................2
Section 2 Setting up the Dodeca Cell for Electrophoresis ......................................3
2.1 Buffer Preparation ..................................................................................................3
2.2 Gel Preparation.......................................................................................................3
2.3 Assembly ...............................................................................................................4
2.4 Required Accessories for Cooling ..........................................................................6
2.5 Running the Gels....................................................................................................6
Section 3 Maintenance..............................................................................................6
Section 4 Troubleshooting Guide.............................................................................7
Section 5 Product Information and Accessories .....................................................8
Section 6 Warranty Information............................................................................11
Section 1
General Information
1.1 Introduction
The Mini-PROTEAN 3 Dodeca Cell is a multi-cell for mini-vertical gel electrophoresis.
It can run four to twelve identical polyacrylamide gels simultaneously. The Dodeca cell
includes clamping frames, buffer dams, a drain line, and gel releasers. Ready Gel precast gels
are compatible with the Mini-PROTEAN 3 Dodeca Cell. Handcasting electrophoresis gels is
also an option; see Section 5 for recommended handcast accessories.
Specifications
Tank and lid Acrylic
Clamping frame Polycarbonate and liquid crystal polymer
Upper electrode holder Polycarbonate with 109 mm (43”) platinum wire
Lower electrode assembly Polycarbonate with 89 cm (35”) platinum wire
Drain line Tygon tubing
Drain line connectors Delrin
Cooling coil Acrylic
Cooling coil connector tubing Tygon
Maximum buffer volume 4.4 liters
Minimum buffer volume 3.4 liters
Overall size 41.5 x 15 x 16.2 cm (L x W x H)
Precast gel compatibility Ready Gel precast gels
Handcast gel compatibility Mini-PROTEAN 3 glass plates and spacers
Safety limits 500V, 500W
Weight 5 kg (11 lb)
Note: Dodeca cell components are not compatible with acetone or ethanol. Use of organic
solvents voids all warranties. The Dodeca cell is compatible only with aqueous reagents.
1.2 Safety
Power to the Mini-PROTEAN 3 Dodeca cell is supplied by an external DC voltage power
supply (not included). The output of the power supply must be isolated from external ground
to insure that the DC voltage output floats with respect to ground. All Bio-Rad power supplies
meet this important safety requirement. Regardless of the power supply used, the maximum
specified operating parameters for the Mini-PROTEAN 3 Dodeca cell are as follows:
500 VDC maximum voltage limit
500 W maximum power limit
40 °C maximum ambient temperature limit
The current to the cell enters the unit through the lid assembly and provides a safety
interlock to the user. The circuit between the cell and the lid’s upper electrodes is broken
when the lid is removed. Always turn off the power supply before removing the lid. Do not
attempt to use the cell without the safety lid.
1
2
Important: This Bio-Rad product is designed and certified to meet *IEC61010-1 and
EN61010-1 safety standards. Certified products are safe to use when operated in accordance
with the instruction manual. This instrument should not be modified or altered in any way.
Alteration of this instrument will
• Void the warranty
• Void the IEC61010-1 and EN61010-1 certifications, and
• Create a potential safety hazard
Bio-Rad is not responsible for any injury or damage caused by use of this instrument for
purposes other than those for which it is intended or by modifications of the instrument not
performed by Bio-Rad or an authorized agent.
*IEC61010-1 and EN61010-1 are internationally accepted electrical safety standards for laboratory instruments.
1.3 Components
To get the best performance from your Mini-PROTEAN 3 Dodeca cell, familiarize
yourself with the components by assembling and disassembling the cell before using it.
Buffer tank and lid The buffer tank and lid combine to fully enclose
the inner chamber during electrophoresis. The
lid cannot be removed without disrupting the
electrical circuit.
Cooling coil The integrated cooling coil chills the lower
buffer chamber. The cooling core must be
connected to a refrigerated circulator.
Electrode assembly The lower buffer chamber contains the anode.
The lid of the Dodeca cell houses the cathode.
Drain port The drain port allows buffer to be removed
from the tank. The drain line attaches via a
quick connect fitting; this assembly is
provided with the cell.
Clamping frame
1
The clamping frame holds two gel sandwiches
in place. It houses the notched gasket.
Buffer dam The molded, one piece buffer dam is used
when running only one gel in a clamping
frame.
Gel releaser The gel releaser is a small green tool that
loosens the two glass plates following
electrophoresis.
Gel sandwich A spacer plate and a short plate with a
polymerized gel form a gel sandwich.
Gel cassette A Ready Gel precast gel or a handcast gel in
an empty Ready Gel cassette is referred to as a
gel cassette.
1
U.S. Patent 6,436,262.
Section 2
Setting up the Dodeca Cell for Electrophoresis
2.1 Buffer preparation
Prepare 3.4–4.4 liters of running buffer (minimum/maximum) for electrophoresis. Buffer
should be equilibrated to room temperature prior to electrophoresis. Premixed buffers are
available (see Section 5).
2.2 Gel Preparation
Ready Gel precast gels or handcast gels using Mini-PROTEAN 3 glass plates can be run
in the Mini-PROTEAN 3 Dodeca cell. Note: All gels in a single electrophoresis experiment
must be of the same gel type, with the same buffer and the same % acrylamide.
2.2.1 Ready Gel Precast Gel Cassette Preparation
Note: The Mini-PROTEAN 3 Dodeca cell is guaranteed only for use with Ready Gel
precast gels.
For complete Ready Gel precast gel instructions, order catalog number 161-0993, Ready Gel
Applications Guide. The Applications Guide is also available on the internet at www.bio-rad.com
in the literature section.
a. Take the Ready Gel precast gel out of its storage pouch.
b. With a razor blade, cut the tape along the black line at the bottom of the gel.
c. Pull the clear tape at the bottom of the Ready Gel Cassette to expose the bottom edge of
the gel.
Repeat for additional Ready Gel precast gels up to a maximum of 12 gels.
Note: If an odd number of gels is to be run, use the buffer dam on the opposite side of the
clamping frame with only one gel to create the clamp assembly.
2.2.2 Handcast Gel Preparation Using Mini-PROTEAN 3 Glass Plates
a. Cast gels per the instructions with the Mini-PROTEAN 3 multi-casting chamber.
b. Remove the gels from the casting chamber or from the storage container.
3
2.3 Assembly
a. Set the clamping frame to the open position.
b. Place gel sandwich or gel cassette into the clamping frame with the Mini-PROTEAN 3
short plates facing inward. Note: the clamping frame requires 2 gels to create the clamp
assembly. If an odd number of gels are being run, use the buffer dam.
c. Lift the gel sandwiches or cassettes into place against the green gaskets. Make sure the
Mini-PROTEAN 3 short plates sit just below the notch in the gasket.
4
d. While gently squeezing the gel sandwiches or cassettes against the green gaskets, slide the
arms of the clamping frame over the gels, locking them into place. This forms the clamp
assembly.
e. The arms of the clamping frame push the short plates up against the notch in the green
gasket, creating a leak-proof seal. At this point, the sample wells can be washed out with
running buffer, and sample can be loaded.
f. Place the Dodeca cell on a stir plate.
g. Lower each clamp assembly into one of the six slots in the tank.
h. Fill the tank with at least 2.5 liters of buffer (3.5 liters recommended) and drop a 38 mm
(1.5") stir bar into the buffer tank. Do not exceed the maximum fill line. Fill each clamp
assembly with 150 ml of running buffer (900 ml total for 6 clamp assemblies.)
i. Turn on the stir plate and make sure the stir bar is mixing effectively.
5
2.4 Required Accessories for Cooling
The Mini-PROTEAN 3 Dodeca cell has a cooling coil with quick connect fittings that
must be connected to a refrigerated circulator. The refrigerated circulator must be able
maintain the buffer temperature of 18–20 ºC during electrophoresis2. Tubing with 3/8" ID
connects the Dodeca cell to the refrigerated circulator. To connect the Dodeca cell to the
refrigerated circulator,
a. Insert the male fittings (supplied in a separate bag) into the tubing on the refrigerated
circulator inlet and outlet tubing.
b. Connect the male fittings from the refrigerated circulator to the female fittings on the
Mini-PROTEAN 3 Dodeca cell. The cooling coil in the Dodeca cell has no specific
orientation, so it is not important which direction the flow goes.
In addition to the refrigerated circulator, the Dodeca cell should be used with a stir bar and
stir plate.
By using the maximum volume of buffer, the refrigerated circulator, and the stir plate, the
effects of heat on electrophoresis results are minimized.
2
The refrigerated circulator used during development of the Dodeca cell was set to 15 ºC and had a flow rate of 3.8 liters per minute.
Under these conditions and using the maximum volume of buffer, the buffer temperature during electrophoresis was maintained
at 18–20 ºC.
2.5 Running the Gels
Insert the red and black color-coded power cables into the corresponding red and black
jacks of the power supply. Set the power supply to run at 200 V constant for 35 minutes. For
best results, use the PowerPac 200 power supply.
Application note:
The initial current draw for one 1.0 mm Ready Gel precast gel in a Laemmli formulation is
about 50 mA; a Dodeca cell filled with 12 precast gels will draw 600 mA at the start of a run.
The current will drop approximately 50% during the run.
Section 3
Maintenance
Mini-PROTEAN 3 Dodeca cell tank and lid, Rinse thoroughly with distilled water
clamping frames, buffer dam, gel releaser after every use.
Glass plates and combs Wash with a laboratory detergent, then
rinse thoroughly with distilled water.
Glass plates and combs (when more stringent Soak in 10 N KOH for 30 minutes (max.)
cleaning is required) and then rinse thoroughly with distilled
water.
The buffer in the lower tank may be reused up to 10 times. Sodium azide (final concentration
of 0.02%) is recommended to help minimize contamination.
Ultimately the number of times the buffer is used depends on the number of gels run each
time and the conditions at which the runs are performed. (The primary concern is ion
depletion.) Prepare fresh buffer if the following problems appear:
• changes in protein mobility
• decrease in band sharpness
• longer run times
• a band at the bottom of the gel which is difficult or impossible to destain
6
Section 4
Troubleshooting Guide
Problem Cause Solution
1. Smile effect – band pattern a. Center of the gel running a. Improper cooling.
curves upward at both sides of hotter than either end Use cooling coil and
the gel refrigerated circulator
to maintain buffer
temperature of 20 ºC.
b. Power conditions are b. Decrease voltage from
excessive 200 V to 150 V or fill
buffer to maximum
volume.
2. Run takes unusually long a. Ion depletion in running a. Prepare and use fresh
time. buffer (lower tank) buffer.
b. Running buffer too b. Check buffer protocol
concentrated in the and dilute buffer if
clamp assembly necessary.
3. Changes in protein mobility a. Ion depletion in running a. Prepare and use fresh
or band sharpness. buffer (lower tank) buffer.
4. Clamp assembly leaks a. Improper assembly a. Be sure the green
gasket is clean, free of
cuts, and is inserted
completely into the
core.
b. Be sure the gel
sandwich or gel
cassette is seated at the
bottom of the clamping
frame before closing
the clamps.
5. SDS is precipitating. a. Running buffer is too cold. a. Set the refrigerated
circulator to maintain a
buffer temperature of
18–20 ºC.
6. Vertical streaking of protein a. Sample overload or a. Dilute sample,
precipitation selectively remove
predominant protein in
the sample, or reduce
voltage by about 25%
to minimize streaking.
b. The ratio of SDS to
protein should be
enough to coat each
protein molecule,
generally 1.4:1. It may
require more SDS for
some membrane
protein samples.
7. Lateral band spreading a. Diffusion out of the wells a. Minimize the time
prior to turning on the between sample
current application and power
start up.
7
Problem Cause Solution
b. Ionic strength of sample b. Use same buffer in
lower than that of gel sample as in gel or
stacking gel.
8. Skewed or distorted bands a. Poor polymerization a. Degas stacking gel
around the sample wells solution thoroughly
prior to casting;
increase ammonium
persulfate and TEMED
concentration by 25%,
wipe comb with
TEMED just before
casting the stacking gel,
insert comb carefully to
prevent trapping of air
bubbles in sample well.
b. Salts in sample. b. Remove salts by
dialysis, desalting
column, etc.
c. Uneven gel interface c. Decrease the
polymerization rate.
Overlay gels very
carefully.
9. Lanes constricted at the a. Ionic strength of sample a. Desalt sample and
bottom of the gel. higher than that of neighboring samples.
surrounding gel.
Section 5
Product Information and Accessories
Catalog
Number Description
Mini-PROTEAN 3 Dodeca Cell and Accessories
165-4100 Mini-PROTEAN 3 Dodeca Cell, includes 6 clamping frames,
2 buffer dams, drain line, and 2 gel releasers
165-4101 Mini-PROTEAN 3 Dodeca Cell, with multi-casting chamber, (includes
165-4100 and 165-4110, order glass plates and combs separately)
165-4102 Replacement clamping frame, 1
165-4103 Lower electrode assembly with platinum wire
165-4104 Replacement drain line
165-4105 Replacement cooling coil, includes connector tubing
165-2948 Replacement power cables
165-3130 Buffer dam
8
Catalog
Number Description
Mini-PROTEAN 3 Multi-Casting Chamber
165-4110 Mini-PROTEAN 3 multi-casting chamber*, includes acrylic blocks and
separation sheets, tapered luer fitting, and stopcock valve
165-4114 Acrylic blocks, 6 mm thickness, 8
165-4115 Separation sheets, 15
165-4116 Mini-PROTEAN 3 multi-casting chamber, with glass plates**,
0.5 mm, includes 15 sets of glass plates
165-4111 Mini-PROTEAN 3 multi-casting chamber, with glass plates**,
0.75 mm, includes 15 sets of glass plates
165-4112 Mini-PROTEAN 3 multi-casting chamber, with glass plates**,
1.0 mm, includes 15 sets of glass plates
165-4113 Mini-PROTEAN 3 multi-casting chamber, with glass plates**,
1.5 mm, includes 15 sets of glass plates
*
Note: Mini-PROTEAN 3 glass plates and combs must be ordered separately
**
Note: Mini-PROTEAN 3 combs must be ordered separately
Mini-PROTEAN 3 Glass Plates and Combs
165-3308 Short plates, 5
165-3309 Spacer plates with 0.5 mm spacers, 5
165-3310 Spacer plates with 0.75 mm spacers, 5
165-3311 Spacer plates with 1.0 mm spacers, 5
165-3312 Spacer plates with 1.5 mm spacers, 5
Combs (2/Pack)
0.5 mm 0.75 mm 1.0 mm 1.5 mm
5-well Comb – 165-3352 165-3357 165-3363
9-well Comb – 165-3353 165-3358 165-3364
10-well Comb 165-3350 165-3354 165-3359 165-3365
15-well Comb 165-3351 165-3355 165-3360 165-3366
Prep/2-D Comb – 165-3356 165-3361 165-3367
IPG Comb – – 165-3362 165-3368
9
Catalog
Number Description
Model 485 Gradient Former
165-4120 Model 485 Gradient Former, includes body with valve stem and tubing con-
nection kit
Power Supply
164-5052 PowerPac™ HC power supply, 100–120/220–240 V
Premixed electrophoresis buffers
161-0772 10x Tris/Glycine/SDS, 5 L
161-0757 10x Tris/Glycine, 5 L
161-0760 10x Tris/Tricine/SDS, 6 x 1 L
161-0770 10x Tris/Boric Acid/EDTA, 5 L
161-0773 50x Tris/Acetic Acid/EDTA, 5 L
10
Section 6
Warranty Information
The Mini-PROTEAN 3 Dodeca cell is warranted for 1 year against defects in materials
and workmanship. If any defects should occur during this warranty period, Bio-Rad
Laboratories will replace the defective parts without charge. However, the following defects
are specifically excluded:
• Defects caused by improper operation
• Repairs or modifications performed by anyone other than Bio-Rad Laboratories or their
authorized agent.
• Damage caused by accidental misuse
• Damage caused by disaster
• Common replacement parts including platinum wire and power cables
• Damage caused by the use of organic solvents
For inquiries or to request repair service, contact your local Bio-Rad office
Warranty Information
Model
Catalog Number
Date of Delivery
Serial Number
Invoice Number
Purchase Order Number
11
4006191 Rev C
Loading...