Bio-Rad Model 491 Prep Cell and Mini Prep Cell - Manual (Page 4)

Table of Contents
Section 1 General Information .................................................................................1
1.1 Introduction ................................................................................................1
1.2 Accessory Equipment ................................................................................1
1.3 Specifications.............................................................................................2
1.4 Chemical Compatability .............................................................................2
1.5 Safety ........................................................................................................2
Section 2 Description of Major Components ..........................................................4
2.1 Model 491 Prep Cell Components .............................................................4
2.2 Lower and Upper Buffer Chamber .............................................................5
2.3 Cooling Core ..............................................................................................5
2.4 Gel Tube Assembly ....................................................................................6
2.5 Elution Chamber Base ...............................................................................6
2.6 Casting Stand ............................................................................................7
Section 3 Assembly and Operation .........................................................................8
3.1 Casting the Preparative Gel ........................................................................8
3.2 Preparing the Frits and Dialysis Membrane ...............................................10
3.3 Assembly of the Elution Chamber ............................................................10
3.4 Assembly of Upper and Lower Buffer Chambers ......................................12
3.5 Purge the Elution Chamber of Air Bubbles ...............................................14
3.6 Cooling the Gel ........................................................................................15
3.7 Loading the Sample .................................................................................16
3.8 Elution Rate, Detection, Collection, and Analysis ......................................16
Section 4 Optimizing Running Conditions for Preparative SDS-PAGE ................17
4.1 Gel Pore Size ...........................................................................................17
4.2 Optimization Procedures ..........................................................................18
4.3 Gel Length Determination for Preparative SDS-PAGE ...............................19
4.4 Gel Tube Size Selection ............................................................................19
4.5 Running Conditions ..................................................................................21
4.6 Elution Times for Proteins .........................................................................21
4.7 Examples of the SDS-PAGE Optimization Procedure ...............................22
Section 5 Disassembling and Cleaning .................................................................26
Section 6 Troubleshooting Guide ..........................................................................27
Section 7 Preparation of Electrophoresis Buffers and Acrylamide
Stock Solutions for SDS-PAGE .............................................................28
7.1 Separating (Resolving) Gel Buffer Stock ...................................................28
7.2 Stacking Gel Buffer Stock ........................................................................28
7.3 Sample Buffer (SDS-Reducing Buffer) Stock ............................................28
7.4 10x Electrode (Running) Buffer, pH 8.3 .....................................................29
7.5 30% Acrylamide Stock Solution ...............................................................29
Section 8 SDS-PAGE Gel Preparation ...................................................................30
8.1 Gel Recipes .............................................................................................30
8.2 Analytical Separating Gel..........................................................................31
8.3 Analytical Stacking Gel .............................................................................31
8.4 Preparative SDS-PAGE Separating and Stacking Gels ............................32