Isotopic Tracer Techniques in Preclinical Research

Description

This book serves as a practical guide and a manual on the application of isotopic tracers in understanding the cellular processes (e.g. molecular interactions, protein synthesis) in growth, development, and disease conditions. The techniques described in the book have been carefully chosen to underscore the role and relevance of isotopic tracers in the identification of molecular targets and mechanisms, and preclinical validation of potential therapeutics. This book is intended to be a valuable resource to non-experts that may include students, researchers, educators/teachers, and others who have an interest to understand the application of isotopic tracers in research. Each chapter includes a background, an outline of the method and underlying principles, and a detailed step-wise protocol. To augment the clarity and intellectual grasp, the chapters also include relevant illustrations and study questions. More importantly, the book covers techniques involving radioactive as well as non-radioactive tracers to expand the scope of isotopes in preclinical research including basic and applied science. Preface Acknowledgement Contents About the Author Chapter 1: Introduction 1.1 Historical Background of Isotopes 1.2 Types of Isotopes and Tracer Techniques 1.2.1 Radioactive Isotopes 1.2.2 Stable Isotopes 1.3 Overview of the General Applications of Isotopic Tracers in Basic and Preclinical Research Chapter 2: Tracer Technique in Basic Research 2.1 Introduction 2.2 Nucleic Acids Interaction: Southern Blotting 2.2.1 Background 2.2.2 Method 2.2.3 Principle 2.2.4 Reagents/Equipment 2.2.5 Stepwise Protocol 2.2.6 Study Questions 2.3 Nucleic Acids Interaction: Northern Blotting 2.3.1 Background 2.3.2 Method 2.3.3 Principle 2.3.4 Reagents/Equipment 2.3.5 Stepwise Protocol 2.3.6 Study Questions 2.4 DNA-Protein Interaction 2.4.1 Background 2.4.2 Method 2.4.3 Principle 2.4.4 Reagents/Equipment 2.4.5 Stepwise Protocol 2.4.6 Study Questions 2.5 Protein Synthesis 2.5.1 Background 2.5.2 Method 2.5.3 Principle 2.5.4 Reagents/Equipment 2.5.5 Stepwise Protocol 2.5.6 Study Questions 2.6 Stable Isotopes in Protein Analysis (SILAC) 2.6.1 Stable Isotope Labeling with Amino Acids in Cell Culture (SILAC) 2.6.2 Background 2.6.3 Method 2.6.4 Principle 2.6.5 Reagents/Equipment 2.6.6 Stepwise Protocol 2.6.7 Study Questions 2.7 Stable Isotopes in Protein Analysis (iTRAQ) 2.7.1 Isobaric Tags for Relative and Absolute Quantitation (iTRAQ) 2.7.2 Background 2.7.3 Method 2.7.4 Principle 2.7.5 Reagents/Equipment 2.7.6 Stepwise Protocol 2.7.7 Study Questions Chapter 3: Tracer Technique in Applied Research 3.1 Introduction 3.2 Pharmacokinetics 3.2.1 Background 3.2.2 Method 3.2.3 Principle 3.2.4 Reagents/Equipment 3.2.5 Stepwise Protocol 3.2.5.1 Sample Collection/Preparation Blood Collection and Fractionation Bladder Puncture-Urine Collection Tissue Harvest and Fixation Tissue Snap Freezing 3.2.5.2 Quantitation of Radioisotope Scintillation Counting 3.2.5.3 Qualitative Analysis of Radioisotope Autoradiography Gel Autoradiography Immunoblotting and LC-MS/MS Tissue Autoradiography 3.2.6 Study Questions 3.3 Pharmacodynamics 3.3.1 Background 3.3.2 Method 3.3.3 Principle 3.3.4 Reagents/Equipment 3.3.5 Stepwise Protocol 3.3.5.1 Imageable Tumor Model 3.3.5.2 Target Validation 3.3.6 Study Questions References Further Reading Index