Robert Hnasko

ELISA: Methods and Protocols - Methods in Molecular Biology Robert Hnasko 2015 edition

Jacket Description/Back: This volume is a practical biochemical guide to the Enzyme-Linked Immunosorbent Assay (ELISA), used to detect a target substance in a liquid sample. The ELISA is an important and widely used diagnostic tool in medicine, animal health, botany and quality assurance processes in food and beverage production. An introductory chapter orients the reader on the basic structure and function of immunoglobulins and their fragments while subsequent chapters outline the methodology to generate monoclonal antibodies using hybridoma technology and the general methods used to purify antibodies. Multiple chapters demonstrate how to creatively use the properties of the antibody to identify, localize and quantify target analytes to answer questions and resolve problems. The reader will learn how to use a variety of immunoassay strategies, reporters and detection systems that will undoubtedly facilitate their efforts to gain answers to their own questions. Written in the 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 protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and easily accessible, "ELISA: Methods and Protocols "seeks to provide both professionals and novices with the technical information necessary for the reader to successfully use the immunoassay as part of the discovery process."Table of Contents:1. The Biochemical Properties of Antibodies and their FragmentsRobert M. Hnasko 2. Hybridoma TechnologyRobert M. Hnasko and Larry H. Stanker 3. Affinity Purification of AntibodiesRobert M. Hnasko and Jeffery A. McGarvey 4. Bio conjugation of Antibodies to Horseradish Peroxidase (HRP) Robert M. Hnasko 5. Indirect ELISAAlice V. Lin 6. Direct ELISAAlice V. Lin 7. A Double-Sandwich ELISA for Identification of Monoclonal Antibodies Suitable for Sandwich ImmunoassaysLarry H. Stanker and Robert M. Hnasko 8. ELISpot and DC-ELISspot Assay to Measure Frequency of Antigen-Specific IFNg Secreting CellsMarcelo A. Navarrete 9. The Western BlotThomas S. Hnasko and Robert M. Hnasko 10. Development of Flow Through Assay for Detection of Antibodies to "Taenia solium" Using Protein-A Colloidal Gold Conjugate as a ProbeChennuru Sreedevi and Panduranga Rao Pavuluri 11. Multiplexed Microsphere Suspension Array-Based Immunoassays Andrew Lin, Alexandra Salvador, and J. Mark Carter 12. Multiplex Immunoassay: A Planar Array on a Chip using the MagArray TechnologyLaurie M. Clotilde, Heng Yu, and M. Luis Carbonell 13. Lateral Flow ImmunoassayKathryn H. Ching 14. Immuno-PCR Assay for Sensitive Detection of Proteins in Real TimeXiaohua He and Stephanie A. Patfield 15. "In-situ" Proximity Ligation Assay (PLA) Sonchita Bagchi, Robert Fredriksson, and Asa Wallen-Mackenzie 16. Tyramide Signal Amplification for Immunofluorescent EnhancementLauren Faget and Thomas S. Hnasko 17. Correlative microscopy for Localization of Proteins In Situ: Pre-Embedding Immuno-Electron Microscopy using FluoroNanogold, Gold Enhancement and Low-Temperature ResinDaniela Boassa 18. Multiplex ELISA using Oligonucleotide Tethered AntibodiesRobert S. Matson 19. Gas Plasma Surface Chemistry for Biological AssaysKhoren Sahagian and Mikki Larner"Publisher Marketing: This volume is a practical biochemical guide to the Enzyme-Linked Immunosorbent Assay (ELISA), used to detect a target substance in a liquid sample. The ELISA is an important and widely used diagnostic tool in medicine, animal health, botany and quality assurance processes in food and beverage production. An introductory chapter orients the reader on the basic structure and function of immunoglobulins and their fragments while subsequent chapters outline the methodology to generate monoclonal antibodies using hybridoma technology and the general methods used to purify antibodies. Multiple chapters demonstrate how to creatively use the properties of the antibody to identify, localize and quantify target analytes to answer questions and resolve problems. The reader will learn how to use a variety of immunoassay strategies, reporters and detection systems that will undoubtedly facilitate their efforts to gain answers to their own questions. Written in the 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 protocols and notes on troubleshooting and avoiding known pitfalls. Authoritative and easily accessible, "ELISA: Methods and Protocols "seeks to provide both professionals and novices with the technical information necessary for the reader to successfully use the immunoassay as part of the discovery process."