Bio-Rad 225 Tube Gel Casting Stand User Manual
Mini-PROTEAN®2-D
Electrophoresis Cell,
Mini-PROTEAN Tube Cell, and
Mini-PROTEAN Tube Module
Instruction
Manual
Catalog Numbers
165-2960,
165-2961 and 165-2965
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 Introduction......................................................................................1
Section 2 Equipment and Reagents...............................................................1
2.1 Equipment ....................................................................................................1
2.2 Description of Major Parts............................................................................2
2.3 Additional Required Equipment ...................................................................3
2.4 Accessories for the Mini Electrophoresis System........................................3
Section 3 Solutions ..........................................................................................4
3.1 Solutions for the First-Dimension.................................................................4
3.2 Preparation of First-Dimension Tube Gels ..................................................5
3.3 Solutions for the Second-Dimension ...........................................................5
Section 4 Safety Instructions..........................................................................6
Section 5 The First-Dimension (IEF) ..............................................................7
5.1 First-Dimension Tube Gels ..........................................................................7
5.2 Casting Tube Gels .......................................................................................7
Section 6 Running the First-Dimension (IEF) ...............................................9
6.1 Electrode Preparation ..................................................................................9
6.2 Sample Preparation ...................................................................................10
6.3 First-Dimension Sample Loading...............................................................10
6.4 Power Conditions .......................................................................................10
Section 7 The Second-Dimension ................................................................11
7.1 Preparation of the Stacking Gel .................................................................11
7.2 Removing the Gel from the Tube and Loading onto the Slab Gel ............11
7.3 Running the Slab Gels ...............................................................................12
Section 8 Maintenance of Equipment ..........................................................12
Section 9 Troubleshooting ............................................................................12
Appendix Electrophoresis Chemicals..........................................................15
Note
To insure best performance from the Mini-PROTEAN 2-D electrophoresis cell, become
fully acquainted with these operating instructions before using the cell to separate samples.
Bio-Rad recommends that you first read these instructions carefully. Then assemble and
disassemble the cell completely without casting a gel. After these preliminary steps, you
should be ready to cast and run a gel.
Bio-Rad also recommends that all Mini-PROTEAN 2-D cell components and accessories
be cleaned with a suitable laboratory cleaner (such as Bio-Rad Cleaning Concentrate,
catalog number 161-0722) and rinsed thoroughly with distilled water, before use.
Warranty
Bio-Rad Laboratories warrants the Mini-PROTEAN 2-D cell against defects in materials
and workmanship for 1 year. If any defects occur in the instrument during this warranty
period, Bio-Rad Laboratories will repair or replace the defective parts free. The following
defects, however, are specifically excluded:
1. Defects caused by improper operation.
2. Repair or modification done by anyone other than Bio-Rad Laboratories or an
authorized agent.
3. Use of fittings or other spare parts supplied by anyone other than Bio-Rad
Laboratories.
4. Damage caused by accident or misuse.
5. Damage caused by disaster.
6. Corrosion due to use of improper solvent or sample.
This warranty does not apply to parts listed below:
1. Platinum wire, glass plates.
For any inquiry or request for repair service, contact Bio-Rad Laboratories after confirming
the model and serial number of your instrument.
Model
Catalog No.
Date of Delivery
Serial No.
Invoice No.
Purchase Order No.
Section 1
Introduction
The benefits of performing 2-D electrophoresis in a mini-format lie primarily in
the reduced time required to complete the entire run. The IEF dimension can be
completed in 2-4 hours. The equilibration step is eliminated or minimal, and the
slab run takes only 35-45 minutes. Using Bio-Rad's protein stains (Bio-Safe
Coomassie, SYPRO Ruby protein stain, Silver stain), the whole 2-D experiment
can be completed in 1 day. Furthermore, the amount of reagents and sample used
is significantly reduced.
This manual describes the procedure for preparing and running the O'Farrell
first-dimension tube gels. For tips on sample preparation, and for alternative gel
and buffer reagent preparation, refer to the FAQs available in the technical support
section of our web site (http://discover.bio-rad.com). Refer to the Mini-PROTEAN
slab cell instruction manual for details on performing the second-dimension
SDS-PAGE electrophoresis.
Section 2
Equipment and Reagents
2.1 Equipment
Catalog
Number Product Description
165-2960 Mini-PROTEAN 2-D Cell, includes tube cell module, 16 sample
reservoirs and stoppers, 50 sample reservoir/capillary tube
connectors, 200 capillary tubes with casting tube, tube gel ejector,
Mini-PROTEAN electrophoresis module, 2 clamp assemblies,
10 sets of glass plates with 1.0 mm spacers, 2-D combs with
1 standard well (2), casting stand with gaskets, alignment card,
leveling bubble, lid with power cables, and instructions.
165-2961 Mini-PROTEAN Tube Cell, includes tube cell module, 16 sample
reservoirs and stoppers, 50 sample reservoir/capillary tube connectors, 200 capillary tubes with casting tube, tube gel ejector,
lower buffer chamber, lid and instructions.
165-2965 Mini-PROTEAN Tube Cell Module, same as 165-2961 without
lower buffer chamber and lid.
Warning:
Do not use alcohol to clean the plastic parts of the Mini-PROTEAN cell. This
will cause damage to the acrylic components. Brief exposure to 10% ethanol
at room temperature is exceptable. If you have questions about a specific
chemical compatibility, please call Bio-Rad’s technical support team (1-8004BIORAD)
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Mini-PROTEAN tube cell (catalog # 165-2961). Includes tube cell module (1), sample reservoirs (2),
capillary tube connectors (3), tube setup; sample reservoirs/capillary tube connector/capillary tube (4),
capillary tubes (5), tube gel ejector (6), and casting tube (7).
2.2 Description of Major Parts
The Mini-PROTEAN 2-D cell (catalog number 165-2960) consists of the following
major parts:
1. Mini-PROTEAN electrophoresis cell for slab gels - A separate manual for the
Mini-PROTEAN cell is included. Refer to it for a complete description of the
slab cell parts.
2. Tube cell module - The tube cell module serves as the upper buffer chamber
and holds up to 16 tubes. The module comes with 16 plastic stoppers to plug
any tube holes not in use.
3. Sample reservoirs - The plastic sample reservoirs are molded with Luer
tapered ends which fit tightly into the 16 tube holes in the tube cell module. Gel
tubes are attached to the reservoirs with flexible Tygon tubing connectors.
4. Flexible tubing connectors (50) - These connectors are small pieces of Tygon
tubing about 7.5 mm long and 2 mm in outer diameter. They are used to connect
the sample reservoirs to the capillary gel tubes.
5. Capillary tubes (200) - The glass capillary tubes are used for running the first
dimension tube gels.
6. Casting tube - The casting tube is about 8.5 cm long and 1.3 cm in diameter. It
is used as a casting chamber to cast up to 40 capillary tube gels at once.
7. Tube gel ejector - The tube gel ejector is a green and white fitting which holds
the glass capillary tubes firmly at one end, without leaking, so that the gels can
be easily extruded. A series of O-rings in the fitting hold the tube tightly when
the gel ejector is used as described. The gel ejector must be used with a 1.0
ml syringe.
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5
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2.3 Additional Required Equipment
The following additional material is required:
1. 1 ml and 10 ml syringes
2. Sample loading syringes, use a 20–100 µl Hamilton syringe because sample
volumes can go up to 100 µl, sample reservoir accommodates a maximum of
200 µl of solution).
3. Intramedic tubing, 0.011" x 0.024" (American Scientific Products catalog number
T6065-1)
4. Parafilm laboratory film
5. Tube gel casting stand (catalog number 165-2020) or ring stand and clamp, to
hold the casting tube vertical
6. Power supply: Bio-Rad's PowerPac 3000 or 1000 power supply are
recommended.
7. Combs and 1.0 mm spacers are needed for the second-dimension. These
parts are supplied with the Mini-PROTEAN 2-D cell (catalog number 165-2960),
but must be purchased separately if the tube cell or tube cell module (catalog
numbers 165-2961 or 165-2965) is being used.
2.4 Accessories for the Mini Electrophoresis System
Catalog
Number Product Description
165-2966 Capillary tubes (200) with casting tube
165-2967 Mini 2-D tube gel ejector
165-2968 Mini-PROTEAN Tube Gel sample reservoirs, 8
165-2969 Mini-PROTEAN Tube module stoppers, 8
165-2970 Mini-PROTEAN Tube module capillary tube connectors, 50
170-3930 Mini Trans-Blot®Electrophoretic Transfer Cell, includes 2 gel
holder cassettes, 4 fiber pads, 1 package of precut blot absorbent
filter paper (thick), modular electrode assembly, Bio-Ice cooling
unit, lower buffer chamber, lid with power cables, instructions.
170-3935 Mini Trans-Blot Module, same as 170-3930 without lower buffer
chamber and lid.
165-2976 Model 422 Electro-Eluter, includes electro-eluter module, 12
Membrane Caps (molecular weight cut off of 12,000-15,000 daltons), 6 Glass Tubes with Frits, 6 Silicone Adaptors, 6 Grommets
and Stoppers, lower buffer chamber with lid and cables, and
instructions.
165-2977 Model 422 Electro-Eluter Module, same as 165-2976 without
lower buffer chamber and lid.
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