Main description:

Progress in functional proteomics has been limited for a long time, partially caused by limitations in assay sensitivity and sample capacity; however, protein microarrays have the ability to overcome these limitations so that a highly parallel analysis of hundreds of proteins in thousands of samples is attainable. In Protein Microarrays: Methods and Protocols, expert researchers in the field present an up-to-date collection of robust strategies in the field of protein microarrays and summarize recent advantages in the field of printing technologies, the development of suitable surface materials, as well as detection and quantification technologies. Written in the highly successful Methods in Molecular Biology (TM) series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and key notes on troubleshooting and avoiding known pitfalls.

Comprehensive and cutting-edge, Protein Microarrays: Methods and Protocols aims to stimulate the application and further advancement of this powerful technology in labs worldwide.

Contents:

Part I: Reverse Phase Protein Arrays

1. Reverse Phase Protein Microarrays for Clinical Applications

Mariaelena Pierobon, Claudio Belluco, and Lance A. Liotta, and Emanuel F. Petricoin III

2. Impact of Blocking and Detection Chemistries on Antibody Performance for Reverse Phase Protein Arrays

Kristi Ambroz

3. Phosphoprotein Stability in Clinical Tissue and Its Relevance for Reverse Phase Protein Microarray Technology

Virginia Espina, Claudius Mueller, and Lance A. Liotta

4. Utilization of RNAi to Validate Antibodies for Reverse Phase Protein Arrays

Heiko Mannsperger, Stefan Uhlmann, Ulrike Korf, and OEzgur Sahin

5. Antibody-Mediated Signal Amplification for Reverse Phase Protein Array-Based Protein Quantification

Jan C. Brase, Heiko Mannsperger, Holger Sultmann, and Ulrike Korf

6. Reverse-Phase Protein Lysate Microarray (RPA) for the Experimental Validation of Quantitative Protein Network Models

Satoshi Nishizuka

7. Characterization of Kinase Inhibitors Using Reverse Phase Protein Arrays

Georg Martiny-Baron, Dorothea Haasen, Daniel D'Dorazio, Johannes Voshol, and Doriano Fabbro

8. Use of Formalin-Fixed and Paraffin-Embedded (FFPE) Tissues for Diagnosis and Therapy in Routine Clinical Settings

Daniela Berg, Katharina Malinowsky, Bilge Ergin, Claudia Wolff, and Karl-Friedrich Becker

9. Producing Reverse Phase Protein Microarrays from Formalin Fixed Tissues

Claudia Wolff, Claudia Schott, Katharina Malinowsky, Daniela Berg, and Karl-Friedrich Becker

10. Use of Reverse Phase Protein Microarrays to Study Protein Expression in Leukemia: Technical and Methodological Lessons Learned

Steven M. Kornblau and Kevin C. Coombes

Part II: Antibody Microarrays

11. Antibody Microarrays as Tools for Biomarker Discovery

Marta Sanchez-Carbayo

12. Assessment of Antibody Specificity Using Suspension Bead Arrays

Jochen M. Schwenk and Peter Nilsson

13. Quantitative Analysis of Phosphoproteins Using Microspot Immunoassays

Frauke Henjes, Frank Goetschel, Anika Joecker, and Ulrike Korf

14. Robust Protein Profiling with Complex Antibody Microarrays in a Dual-Color Mode

Christoph Schroeder, Mohamed S.S. Alhamdani, Kurt Fellenberg, Andrea Bauer, Anette Jacob, and Joerg D. Hoheisel

15. High-Throughput Studies of Protein Glycoforms Using Antibody-Lectin Sandwich Arrays

Brian B. Haab and Tingting Yue

16. Microspot Immunoassay-Based Analysis of Plasma Protein Profiles for Biomarker Discovery Strategies

Johanna Sonntag, Heiko Mannsperger, Anika Joecker, and Ulrike Korf

17. Recombinant Antibodies for the Generation of Antibody Arrays

Carl A.K. Borrebaeck and Christer Wingren

Part III: Protein Microarrays

18. Producing Protein Microarrays from DNA Microarrays

Oda Stoevesandt, Michael J. Taussig, and Mingyue He

19. Cell Arrays and High-Content Screening

Holger Erfle, Anastasia Eskova, Jurgen Reymann, and Vytaute Starkuviene

20. Probing Calmodulin Protein-Protein Interactions Using High Content Protein Arrays

David J. O'Connell, Mikael Bauer, Sara Linse, and Dolores J. Cahill

21. Protein Function Microarrays for Customized Systems-Oriented Proteome Analysis

Jonathan M. Blackburn and Aubrey Shoko

22. Optimized Autoantibody Profiling on Protein Arrays

Sara L. O'Kane, John K. O'Brien, and Dolores J. Cahill

Part IV: Sample Immobilization Strategies

23. Inkjet Printing for the Production of Protein Microarrays

Iain McWilliam, Marisa Chong Kwan, and Duncan Hall

24. Impact of Substrates for Probe Immobilization

Ursula Sauer

25. Contact Printing of Protein Microarrays

John Austin