Handbook of ELISPOT (2^nd Ed., Softcover reprint of the original 2nd ed. 2012) Methods and Protocols Methods in Molecular Biology Series, Vol. 792

Enzyme-linked immunospot assay (ELISPOT) has been known for some time as a unique state-of-the-art technique for studying the cytokine-secreting activity of immune system cells, and it appears to be one of the fast growing applications in biomedical research, becoming an indispensable tool in vaccine development, HIV research, transplantation studies, and cancer and allergy research.  The second edition of Handbook of ELISPOT: Methods and Protocols, only the second book in the field which is entirely dedicated to ELISPOT assay, shares the detailed techniques that have been developed since the release of the popular first edition.  Straight from the labs of seasoned experts, this book covers setting and performing ELISPOT assays, ELISPOT for veterinary research, advanced ELISPOT techniques, image and data analysis, as well as vaccine development and diagnostics.  Written in the highly successful Methods in Molecular Biology? series format, chapters include introductions to their respective chapters, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Unique Strengths of ELISPOT for T Cell Diagnostics.- The Impact of Harmonization on ELISPOT Assay Performance.- Equine ELISPOT Assay to Study Secretion of IFNγ and IL-4 from Peripheral Blood Mononuclear Cells.- Utilization of Feline ELISPOT for Mapping Vaccine Epitopes.- Analyzing Cellular Immunity to AAV in a Canine Model Using ELISPOT Assay.- Dual and Triple Color Fluorospot.- ELISPOT Assay as a Tool to Study Oxidative Stress in Lymphocytes.- ELISPOT Assay for Neuroscience Research: Studying TNFa Secretion from Microglial Cells.- ELISPOT Assay as a Tool to Study the Effects of Stem Cells on Cytokine Secretion.- Combining ELISPOT and ELISA to Measure Amounts of Cytokines Secreted by a Single Cell.- How ELISPOT Morphology Reflects on the Productivity and Kinetics of Cells’ Secretory Activity.- Mathematical Algorithms for Automatic Search, Recognition, and Detection of Spots in ELISPOT Assay.- Objective, User-Independent ELISPOT Data Analysis Based on Scientifically Validated Principles.- Statistical Analysis of ELISPOT Assays.- Response Determination Criteria for ELISPOT: Toward a Standard that Can Be Applied Across Laboratories.- Detection of Vaccinia Virus-Specific IFNγ and IL-10 Secretion from Human PBMCs and CD8⁺ T Cells by ELISPOT.- ELISPOT Assays to Enumerate Bovine IFN-γ Secreting Cells for the Development of Novel Vaccines Against Bovine Tuberculosis.- IL-7 Addition Increases Spot Size and Number as Measured by T-SPOT.TB®.- Overview of Membranes and Membrane Plates Used in Research and Diagnostic ELISPOT Assays.