Receptor Tyrosine Kinases: Methods and Protocols

Jacket Description/Back:"Receptor Tyrosine Kinases: Methods and Protocols" explores on the most widely studied of the protein classes, known for their central role in several cellular functions and in a variety of human pathologies. Beginning with a section on activation and signaling of Receptor Tyrosine Kinases (RTKs), the book continues with chapters on aberrant expression of RTKs, trafficking and negative regulation, as well as RTKs as pharmacological targets. Written in the highly successful "Methods in Molecular Biology" series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, "Receptor Tyrosine Kinases: Methods and Protocols" contains an up-to-date overview of the most relevant and widely used methods employed in this field, providing detailed protocols that molecular and cellular biologists could easily adopt in their research programs on RTKs. Table of Contents: Part I: RTKs Activation and Signalling 1. Analysis of Receptor Tyrosine Kinase (RTK) Phosphorylation by Immunoblotting Martina McDermott and Norma O Donovan 2. Analysis of Changes in Phosphorylation of Receptor Tyrosine Kinases: Antibody Arrays Sweta Rani and Lorraine O Driscoll 3. Analysis of Epidermal Growth Factor Receptor Dimerization by BS3 Crosslinking Harmony F. Turk and Robert S. Chapkin 4. Single-Molecule Optical Methods Analyzing Receptor Tyrosine Kinase Activation in Living Cells Inhee Chung and Ira Mellman 5. Evaluation of the Dimerization Profiles of HER Tyrosine Kinases by Time-Resolved Forster Resonance Energy Transfer (TR-FRET) Evelyne Lopez-Crapez, Alexandre Ho-Pun-Cheung, Patrick Garnero, and Herve Bazin 6. Applying the Proximity Ligation Assay (PLA) to Mouse Preimplantation Embryos for Identifying Protein-Protein Interactions In Situ Ivan Bedzhov and Marc P. Stemmler Part II: RTKs Aberrant Expression 7. Analysis of Receptor Tyrosine Kinase Gene Amplification on the Example of "FGFR1" Diana Boehm, Anne von Massenhausen, and Sven Perner 8. Quantification of the Effects of Mutations on Receptor Tyrosine Kinase (RTK) Activation in Mammalian Cells Lijuan He and Kalina Hristova Part III: RTKs Trafficking and Negative Regulation 9. Cell Surface Biotinylation of Receptor Tyrosine Kinases to Investigate Intracellular Trafficking Mathieu J. F. Crupi, Douglas S. Richardson, and Lois M. Mulligan 10. Studying N-Linked Glycosylation of Receptor Tyrosine Kinases Harri M. Itkonen and Ian G. Mills 11. Identification of Receptor Protein Tyrosine Phosphatases (RPTPs) as Regulators of Receptor Tyrosine Kinases (RTKs) Using an RPTP siRNA-RTK Substrate Screen Hojin Lee and Anton M. Bennett 12. Downregulation of Receptor Tyrosine Kinases Through Ubiquitination: Analysis by Immunodetection Noriaki Shimokawa and Noriyuki Koibuchi 13. Regulation of Receptor Tyrosine Kinases by miRNA: Overexpression of miRNA Using Lentiviral Inducible Expression Vectors XiangDong Le, Andrew T. Huang, Yunyun Chen, and Stephen Y. Lai Part IV: RTKs as Pharmacological Targets 14. Human Tumor Xenografts in Mouse as a Model for Evaluating Therapeutic Efficacy of Monoclonal Antibodies or Antibody-Drug Conjugate Targeting Receptor Tyrosine Kinases Liang Feng, Wei Wang, Hang-Ping Yao, Jianwei Zhou, Ruiwen Zhang, and Ming-Hai Wang 15. Receptor Tyrosine Kinase Targeting in Multicellular Spheroids Susan Breslin and Lorraine O Driscoll 16. Receptor Tyrosine Kinases and Drug Resistance: Development and Characterization of In Vitro Models of Resistance to RTK Inhibitors Claire Corcoran and Lorraine O Driscoll"