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)
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Table of Contents |
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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.
Model
Catalog No.
Date of Delivery
Serial No.
Invoice No.
Purchase Order No.
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.
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 |
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Number |
Product Description |
165-2960 |
Mini-PROTEAN 2-D Cell, includes tube cell module, 16 sample |
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reservoirs and stoppers, 50 sample reservoir/capillary tube |
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connectors, 200 capillary tubes with casting tube, tube gel ejector, |
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Mini-PROTEAN electrophoresis module, 2 clamp assemblies, |
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10 sets of glass plates with 1.0 mm spacers, 2-D combs with |
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1 standard well (2), casting stand with gaskets, alignment card, |
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leveling bubble, lid with power cables, and instructions. |
165-2961 |
Mini-PROTEAN Tube Cell, includes tube cell module, 16 sample |
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reservoirs and stoppers, 50 sample reservoir/capillary tube con- |
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nectors, 200 capillary tubes with casting tube, tube gel ejector, |
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lower buffer chamber, lid and instructions. |
165-2965 |
Mini-PROTEAN Tube Cell Module, same as 165-2961 without |
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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 (1-800- 4BIORAD)
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5
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2 |
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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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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 |
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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 |
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holder cassettes, 4 fiber pads, 1 package of precut blot absorbent |
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filter paper (thick), modular electrode assembly, Bio-Ice cooling |
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unit, lower buffer chamber, lid with power cables, instructions. |
170-3935 |
Mini Trans-Blot Module, same as 170-3930 without lower buffer |
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chamber and lid. |
165-2976 |
Model 422 Electro-Eluter, includes electro-eluter module, 12 |
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Membrane Caps (molecular weight cut off of 12,000-15,000 dal- |
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tons), 6 Glass Tubes with Frits, 6 Silicone Adaptors, 6 Grommets |
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and Stoppers, lower buffer chamber with lid and cables, and |
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instructions. |
165-2977 |
Model 422 Electro-Eluter Module, same as 165-2976 without |
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lower buffer chamber and lid. |
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