Receptor Tyrosine Kinases: Methods and Protocols

ISBN-13: 9781493917884 / Angielski / Twarda / 2014 / 181 str.

Serena Germano

Opis

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.

Spis treści

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 Förster Resonance Energy Transfer (TR-FRET)

            Evelyne Lopez-Crapez, Alexandre Ho-Pun-Cheung, Patrick Garnero, and Hervé 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 Mässenhausen, 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