Main description:

This third edition volume expands on the previous editions with new protocols and up-to-date techniques to help researchers excel in this field. The chapters in this book cover topics such as ELISpot's sibling (FluoroSpot); ELISpot for influenza, Dengue virus, and tuberculosis; and vaccine research, essentials control, and assay evaluation in ELISpot. 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, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Cutting-edge and comprehensive, Handbook of ELISPOT: Methods and Protocols, Third Edition is a valuable resource for both novice researchers and experienced ELISpot users."

Contents:

Preface...Table of Contents...Contributing Authors...
1. Challenges in Developing Protein Secretion Assays at a Single-Cell LevelYoshitaka Shirasaki and Osamu Ohara
2. To Count of Not to Count: ELISPOT Evaluation RevisitedSylvia Janetzki
3. Essential Controls for ELISpot AssayJodi Hagen and Alex Kalyuzhny
4. Automatic Search of Spots and Color Classification in ELISPOT AssaySergey S. Zadorozhny and Nikolai N. Martynov
5. Four Color ImmunoSpot (R) Assays for Identification of Effector T Cell LineagesJodi Hanson, Diana R. Roen, and Paul V. Lehmann
6. Detection of Cross-Reactive B Cells Using the FluoroSpot Assay Peter Jahnmatz and Niklas Ahlborg
7. Multiplex ImmunoSpot (R) Assays for the Study of Functional B Cell SubpopulationsDiana R. Roen, Jodi Hanson, and Paul V. Lehmann
8. Detecting All Immunoglobulin Classes and Subclasses in Multiplex 7 Color ImmunoSpot (R) AssaysRichard Caspell and Paul V. Lehmann
9. Multiplexing T- and B-Cell FLUOROSPOT Assays: Experimental Validation of the Multi-Color ImmunoSpot (R) Software Based on Center of Mass Distance AlgorithmAlexey Y. Karulin, Zoltan Megyesi, Richard Caspell, Jodi Hanson, and Paul V. Lehmann
10. Multi-Color FLUOROSPOT Counting Using ImmunoSpot (R) Fluoro-X (TM) SuiteZoltan Megyesi, Paul V. Lehmann, and Alexey Y. Karulin
11. B-Cell ELISpot Assay to Quantify Antigen-Specific Antibody-Secreting Cells in Human Peripheral Blood Mononuclear Cells Haw Hwai, Yi-Ying Chen, and Siang-Jong Tzeng
12. Identification of Novel Mycobacterial Targets for Murine CD4+T Cells by IFN ELISPOTAlison J. Johnson, Steven C. Kennedy, Tony W. Ng, and Steven A. Porcelli
13. ELISPOT-Based "Multi-Color FluoroSpot" to Study Type-Specific and Cross-Reactive Responses in Memory B Cells after Dengue and Zika Virus InfectionsPaulina Andrade, Josefina Coloma, and Eva Harris
14. Cultured ELISpot Assay to Investigate Dengue Virus Specific T Cell ResponsesChandima Jeewandara, Graham S. Ogg, and Gathsaurie Neelika Malavige
15. Ex Vivo ELISpot Assay to Investigate Dengue Virus Specific T Cell ResponsesGathsaurie Neelika Malavige
16. Ex Vivo ELISpot Assay to Investigate iNKT Cell Responses in Acute Dengue InfectionAchala Indika Kamalasasa and Gathsaurie Neelika Malavige
17. Dendritic Cell-Based ELISpot Assay for Assessing T Cell-IFN- Responses in Human Peripheral Blood Mononuclear Cells to Dengue Envelope ProteinsPeifang Sun and Monika Simmons
18. Utilization of Feline ELISpot to Evaluate the Immunogenicity of a T Cell-Based FIV MAP VaccinesBikash Sahay, Alek M. Aranyos, Andrew McAvoy, and Janet K. Yamamoto
19. Detection and Quantification of Influenza A/H1N1 Virus-Specific Memory B Cells in Human PBMCs using ELISpot AssayIana H. Haralambieva, Inna G. Ovsyannikova, Richard B. Kennedy, and Gregory A. Poland