Synopses & Reviews
About 17 million Americans suffer from diabetes mellitus, a disease caused by defects in insulin production, insulin action, or both, and also a leading cause of kidney failure, blindness, and amputations. In Diabetes Mellitus: Methods and Protocols, leading researchers and clinical investigators describe their cutting-edge techniques for studying these defects at both the molecular and biochemical levels. Written in step-by-step detail to ensure ready reproducibility and robust results, these techniques allow investigators, both novices and those already active in the field, to study every major facet of insulin production and action. Each protocol includes an introduction to the technique, an explanation of its application, and a list of materials. Practical notes discuss how to avoid pitfalls, as well as how to adapt the methods to your own research. Cutting-edge and highly practical, Diabetes Mellitus: Methods and Protocols provides basic scientists and clinical researchers, both experienced investigators and those new to the field, all the critical tools needed for the productive study of diabetes mellitus today.
Review
"This book provides all the critical tools needed for the productive study of diabetes mellitus today." - Clinical Laboratory International "Each chapter/protocol is well referenced and the book is indexed. Because Diabetes Mellitus: Methods and Protocols provides all the critical tools needed for the productive study of diabetes mellitus, it is a necessary purchase for libraries supporting basic research programs that study the molecular and biochemical bases of diabetes mellitus." -E-STREAMS
Synopsis
Diabetes mellitus is the collective name for a group of diseases associated with hyperglycemia (high levels of blood glucose) caused by defects in insulin p- duction, insulin action, or both. About 6. 2% of the US population (17 million people) have diabetes mellitus. It is the leading cause of kidney failure, bli- ness, and amputations. It is also a major risk factor for heart diseases, stroke, and birth defects. Diabetes Mellitus: Methods and Protocols provides a state-of-the-art account of the experimental methodology for studying the molecular defects leading to diabetes mellitus, both at the molecular and biochemical levels. The chapters cover a wide range of topics written by experts in their respective fields and are organized in two sections: Insulin Production and Insulin Action. The detailed experimental protocols presented, including the notes of interest, provide a very useful tool for basic researchers and clinicians for investigating and treating this disease. Each chapter starts with an introduction to a specific technique and explains its application in the field of diabetes research. Following the list of materials, a detailed description of the technique is presented in the methods section in a way that enables the successful execution of the protocol. The "Notes" section at the end discusses the pitfalls of the technique and alternative approaches. I am grateful to the numerous scientists who have contributed to this volume by writing both highly detailed and understandable chapters.
Synopsis
Leading researchers and clinical investigators describe their best cutting-edge techniques for studying, at both the molecular and biochemical levels, the defects in insulin production and action associated with diabetes. Written in step-by-step detail to ensure ready reproducibility and robust results, these techniques allow investigators, both novices and those already active in the field, to study every major facet of insulin production and action. Each protocol includes an introduction to the technique, an explanation of its application, and a list of materials. Practical notes discuss how to avoid pitfalls, as well as how to adapt the methods to your own research.
Table of Contents
Part I. Insulin Production Isolation of Islets of Langerhans from Rodent Pancreas Colleen B. Kelly, Libby A. Blair, John A. Corbett, and Anna L. Scarim Purification of Rat Pancreatic b-Cells by Fluorescence-Activated Cell Sorting Geert Stangé, Mark Van De Casteele, and Harry Heimberg Assessment of Insulin Secretion in the Mouse Marcela Brissova, Wendell E. Nicholson, Masakazu Shiota, and Alvin C. Powers Detection of Insulin Production by Immunohistochemistry Susan C. Campbell and Wendy M. Macfarlane Quantification of the Level of Insulin Gene Expression Susan C. Campbell and Wendy M. Macfarlane Chromatin Immunoprecipitation Using Isolated Islets of Langerhans Marcelina Párrizas, Sylvia F. Boj, Reini F. Luco, Miguel Angel Maestro, and Jorge Ferrer Adenoviral Gene Transfer into b-Cell Lines Amber L. Mosley and Sabire Özcan Utilization of NOD Mice in the Study of Type 1 Diabetes Dennis G. Karounos and Susan E. Goes Part II. Insulin Action Introduction of DNA into 3T3-L1 Adipocytes by Electroporation Shuichi Okada, Masatomo Mori, and Jeffrey E. Pessin Analysis of Insulin-Stimulated Glucose Uptake in Differentiated 3T3-L1 Adipocytes Jaganathan Lakshmanan, Jeffrey S. Elmendorf, and Sabire Özcan Fractionation Analysis of the Subcellular Distribution of GLUT-4 in 3T3-L1 Adipocytes Jeffrey S. Elmendorf Visualization and Quantitation of Integral Membrane Proteins Using a Plasma Membrane Sheet Assay John B. Knight and Ann Louise Olson Assaying Tyrosine Phosphorylation of Insulin Receptor and Insulin Receptor Substrates Eileen L. Whiteman and Morris J. Birnbaum Measuring Insulin-Stimulated Phosphatidyl-Inositol 3-Kinase Activity Li-Ping Wang and Scott A. Summers Assaying AKT/ Protein Kinase B Activity Kyle L. Hoehn and Scott A. Summers Measurements of Cellular Phosphoinositide Levels in 3T3-L1 Adipocytes Daryll B. DeWald Measurement of Glycogen Synthesis and Glycogen Synthase Activity in 3T3-L1 Adipocytes Matthew J. Brady Measurement of Contraction-Stimulated GLUT-4 Translocation in Isolated Skeletal Muscle Joseph T. Brozinick, Jr. Single Embryo Measurement of Basal- and Insulin-Stimulated Glucose Uptake Maggie M-Y. Chi and Kelle H. Moley Immunohistologic Staining of Muscle and Embryos to Detect Insulin-Stimulated Translocation of Glucose Transporters Mary O. Carayannopoulos and Kelle H. Moley Index