Bio-Rad Model 111 Mini IEF Cell User Manual

Page 3

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Table of Contents

Section 1

General Information................................................................................1

1.1

Introduction ...........................................................................................................1

1.2

Specifications.........................................................................................................1

1.3

Safety .....................................................................................................................2

1.4

Model 111 Mini IEF Cell Components .................................................................3

1.5

Capillary Thin Layer Gel Casting Tray .................................................................3

Section 2

Introduction to Isoelectric Focusing.......................................................4

2.1

The Electrofocusing Principle ...............................................................................4

2.2

Carrier Ampholytes ...............................................................................................4

2.3

Choice of Support Matrix ......................................................................................5

Section 3

Polyacrylamide Gel Isoelectric Focusing ...............................................5

3.1

Considerations in Matrix Preparation....................................................................5

3.2

Stock Solutions for Polyacrylamide IEF Gels.......................................................6

3.3

Reagents for Polyacrylamide Electrofocusing Gels ..............................................6

3.4

Use of Gel Support Film for Polyacrylamide........................................................7

3.5

Casting Polyacrylamide Gels ................................................................................8

3.6

Sample Preparation................................................................................................9

3.7

Sample Application .............................................................................................10

3.8

Position of Application........................................................................................10

Section 4

Running The Gel....................................................................................10

4.1

Set Up Proceedure ...............................................................................................10

4.2

Run Conditions ....................................................................................................11

Section 5

Sample Detection and Gel Storage .......................................................12

5.1

Removing the Gel................................................................................................12

5.2

Band Detection ....................................................................................................12

5.3

Destaining............................................................................................................13

5.4

Other Detection Methods ....................................................................................13

5.5

Gel Drying and Preservation ..............................................................................13

Section 6

Agarose Gel Electrofocusing .................................................................14

6.1

Introduction .........................................................................................................14

6.2

Preparing Agarose Gels .......................................................................................14

6.3

Sample Preparation..............................................................................................16

6.4

Sample Application .............................................................................................16

6.5

Position of Application........................................................................................16

6.6

Set Up Procedure .................................................................................................17

6.7

Run Conditions ....................................................................................................17

6.8

Sample Detection.................................................................................................18

Section 7

Troubleshooting .....................................................................................19

7.1

General Troubleshooting .....................................................................................19

7.2

Casting Troubleshooting, Polyacrylamide Gels ..................................................20

7.3

Casting Troubleshooting, Agarose Gels ..............................................................21

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