Library Science

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. Authors: Ozcan, Sabire

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Table of Contents

1. Isolation of Islets of Langerhans from Rodent Pancreas (pp. 003-014)
   Kelly, Colleen B.; Blair, Libby A.; Corbett, John A.; Scarim, Anna L.
2. Purification of Rat Pancreatic ?÷-Cells by Fluorescence-Activated Cell Sorting (pp. 015-022)
   Stange, Geert; Van De Casteele, Mark; Heimberg, Harry
3. Assessment of Insulin Secretion in the Mouse (pp. 023-045)
   Brissova, Marcela; Nicholson, Wendell E.; Shiota, Masakazu; Powers, Alvin C.
4. Detection of Insulin Production by Immunohistochemistry (pp. 047-049)
   Campbell, Susan C.; Macfarlane, Wendy M.
5. Quantification of the Level of Insulin Gene Expression (pp. 051-059)
   Campbell, Susan C.; Macfarlane, Wendy M.
6. Chromatin Immunoprecipitation Using Isolated Islets of Langerhans (pp. 061-071)
   Parrizas, Marcelina; Boj, Sylvia F.; Luco, Reini F.; Maestro, Miguel Angel; Ferrer, Jorge
7. Adenoviral Gene Transfer into ?÷-Cell Lines (pp. 073-079)
   Mosley, Amber L.; Ozcan, Sabire
8. Utilization of NOD Mice in the Study of Type 1 Diabetes (pp. 081-090)
   Karounos, Dennis G.; Goes, Susan E.
9. Introduction of DNA into 3T3-L1 Adipocytes by Electroporation (pp. 093-096)
   Okada, Shuichi; Mori, Masatomo; Pessin, Jeffrey E.
10. Analysis of Insulin-Stimulated Glucose Uptake in Differentiated 3T3-L1 Adipocytes (pp. 097-103)
    Lakshmanan, Jaganathan; Elmendorf, Jeffrey S.; Ozcan, Sabire
11. Fractionation Analysis of the Subcellular Distribution of GLUT-4 in 3T3-L1 Adipocytes (pp. 105-111)
    Elmendorf, Jeffrey S.
12. Visualization and Quantitation of Integral Membrane Proteins Using a Plasma Membrane Sheet Assay (pp. 113-118)
    Knight, John B.; Olson, Ann Louise
13. Assaying Tyrosine Phosphorylation of Insulin Receptor and Insulin Receptor Substrates (pp. 119-126)
    Whiteman, Eileen L.; Birnbaum, Morris J.
14. Measuring Insulin-Stimulated Phosphatidyl-Inositol 3-Kinase Activity (pp. 127-136)
    Wang, Li-Ping; Summers, Scott A.
15. Assaying AKT/Protein Kinase B Activity (pp. 137-144)
    Hoehn, Kyle L.; Summers, Scott A.
16. Measurements of Cellular Phosphoinositide Levels in 3T3-L1 Adipocytes (pp. 145-154)
    DeWald, Daryll B.
17. Measurement of Glycogen Synthesis and Glycogen Synthase Activity in 3T3-L1 Adipocytes (pp. 155-161)
    Brady, Matthew J.
18. Measurement of Contraction-Stimulated GLUT-4 Translocation in Isolated Skeletal Muscle (pp. 163-169)
    Brozinick, Jr., Joseph T.
19. Single-Embryo Measurement of Basal- and Insulin-Stimulated Glucose Uptake (pp. 171-178)
    Chi, Maggie M-Y.; Moley, Kelle H.
20. Immunohistologic Staining of Muscle and Embryos to Detect Insulin-Stimulated Translocation of Glucose Transporters (pp. 179-185)
    Carayannopoulos, Mary O.; Moley, Kelle H.

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