This manual describes the methodology and applications of microinjection, a powerful technique for transposing material into living cells through needle injection. Information is presented on the techniques principles, the required equipment, preparation of receiving cells and injected material, and the analysis of results. Detailed protocols for a wide range of applications in different cellular systems are included. A large part of the book is also dedicated to the Xenopus laevis oocyte system.
This manual describes the methodology and applications of microinjection, a powerful technique for transposing material into living cells through needle injection. Information is presented on the techniques principles, the required equipment, preparation of receiving cells and injected material, and the analysis of results. Detailed protocols for a wide range of applications in different cellular systems are included. A large part of the book is also dedicated to the Xenopus laevis oocyte system.
1. Needle Microinjection: A Brief History.- 2. Microinjection of Macromolecules into Mammalian Cells in Culture.- 3. The Use of Plasmid Microinjection to Study Specific Cell Cycle Phase Transitions.- 4. Combining Microinjection with Microanalytical Methods to Investigate Regulation of Metabolic Pathways.- 5. Import of Stably Folded Proteins into Peroxisomes.- 6. Study of the Function of Microtubule Proteins.- 7. Cytoskeleton Regulation by Rho Proteins.- 8. Cytoskeletal Regulation by Rho-p21 Associated Kinases: Analysis of ROK and PAK Function by Plasmid Microinjection.- 9. Use of Microinjection to Study Apoptosis and its Prevention.- 10. Role of the InsP3 Receptor in Intracellular Ca2+ Release and Ca2+ Entry.- 11. Microinjection of Xenopus laevis Oocytes: A Model System.- 12. Xenopus laevis Oocytes as a Model for Studying the Activation of Intracellular Kinases.- 13. Signal Transduction in Xenopus laevis Oocytes by Ras Oncoproteins and Lipid Metabolites.- 14. The Multiple Roles of Mos During Meiosis.- 15. Use of the Xenopus Oocyte System to Study RNA Transport.- 16. Functional Expression of G Protein-Coupled Receptors in Xenopus laevis Oocytes.- 17. Expression of Glucose Transporters in Xenopus laevis Oocytes: Applications for the Study of Membrane Transporter Function and Regulation.- 18. DNA Injection into Xenopus laevis Embryos as a Tool to Study Spatial Gene Activity.- 19. Transformation of Nematodes by Microinjection.- Guide to Solutions.- Guide to Protocols.- Troubleshooting Guide.
1. Needle Microinjection: A Brief History.- 2. Microinjection of Macromolecules into Mammalian Cells in Culture.- 3. The Use of Plasmid Microinjection to Study Specific Cell Cycle Phase Transitions.- 4. Combining Microinjection with Microanalytical Methods to Investigate Regulation of Metabolic Pathways.- 5. Import of Stably Folded Proteins into Peroxisomes.- 6. Study of the Function of Microtubule Proteins.- 7. Cytoskeleton Regulation by Rho Proteins.- 8. Cytoskeletal Regulation by Rho-p21 Associated Kinases: Analysis of ROK and PAK Function by Plasmid Microinjection.- 9. Use of Microinjection to Study Apoptosis and its Prevention.- 10. Role of the InsP3 Receptor in Intracellular Ca2+ Release and Ca2+ Entry.- 11. Microinjection of Xenopus laevis Oocytes: A Model System.- 12. Xenopus laevis Oocytes as a Model for Studying the Activation of Intracellular Kinases.- 13. Signal Transduction in Xenopus laevis Oocytes by Ras Oncoproteins and Lipid Metabolites.- 14. The Multiple Roles of Mos During Meiosis.- 15. Use of the Xenopus Oocyte System to Study RNA Transport.- 16. Functional Expression of G Protein-Coupled Receptors in Xenopus laevis Oocytes.- 17. Expression of Glucose Transporters in Xenopus laevis Oocytes: Applications for the Study of Membrane Transporter Function and Regulation.- 18. DNA Injection into Xenopus laevis Embryos as a Tool to Study Spatial Gene Activity.- 19. Transformation of Nematodes by Microinjection.- Guide to Solutions.- Guide to Protocols.- Troubleshooting Guide.
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