TABLE OF CONTENTS PREFACE FOR INDIVIDUAL EXPERIMENTS 101-106 The material in this folder is divided into three sections. The first section provides basic information on the biochemistry of proteins and is also intended to acquaint the students with the principles and techniques of electrophoresis. T h e second part shows students how to apply what they have learned to perform one or more exercises i n modern biology. E a c h exercise consists of a background information section. an experimental procedure and study questions. Befor e the laboratory, the student should read the background information section, study the directions for doing the experiment and understand the reason for each step in the procedure. The instructor manual for experiments 101-106 is the third item in the folder. T h i s manual contains a listing of the chemicals provided with each experiment in this series, procedures for preparing solutions, typical experimental results and answers to study questions. John N. Anderson, Ph.D. Professor of Biology Department of Biological Sciences Purdue University West Lafayette, Indiana 47907 Co 1986 by John N. Anderson. All rights reserved.
NOTES: 1. Permission is granted to reproduce the written material in this folder one time for educational purposes only. 2. Electrophoresis Package 1 is required to perform the exercises described in this folder.
PART A. BACKGROUND I NFORMATI ON
Protein Composition and Structure: A Review of the Basics Amino Acids-Building Blocks of Proteins The Peptide Bond Three-Dimensional Protein Structure Theoretical Aspects of Electrophoresis General Description Separation of Amino Acids and Proteins by Electrophoresis Practical Aspects of Electrophoresis Electrophoresis Equipment Electrophoresis Chemicals Accessories IV. Procedures for the Preparation. Electrophoresis and Staining of Agarose Gels Pouring the Agarose Gels Sample Application Electrophoresis Staining and Dcstaining V. Suggested Reading and References for Part A
2 3
6 6 8 10 11
13 14 15 16 16
PART B. LABORATORY EXERCISES Experiment I. Elecm)phoretic Separation of Proteins
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Experiment 2. Sickle Cell Anemia
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Experiment 3. Analysis of Human Serum Proteins
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Experiment 4_ Evolution of Serum Proteins
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Experiment 5. Detection of Amylase
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Experiment 6. Serum Enzymes and the Diagnosis of Disease
40
Suggested Reading and References for Part B
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