Hnasko R. (Ed.) ELISA. Methods and Protocols

Humana Press, 2015 — 216p. — (Methods in Molecular Biology 1318) — ISBN: 978-1-4939-2742-5 (eBook), 978-1-4939-2741-8 (Hardcover).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.The Biochemical Properties of Antibodies and Their Fragments
Hybridoma Technology
Affinity Purification of Antibodies
Bioconjugation of Antibodies to Horseradish Peroxidase (HRP)
Indirect ELISA
Direct ELISA
A Double-Sandwich ELISA for Identification of Monoclonal Antibodies Suitable for Sandwich Immunoassays
ELISpot and DC-ELISpot Assay to Measure Frequency of Antigen-Specific IFNγ-Secreting Cells
The Western Blot
Flow-Through Assay for Detection of Antibodies Using Protein-A Colloidal Gold Conjugate as a Probe
Multiplexed Microsphere Suspension Array-Based Immunoassays
Multiplex Immunoassay: A Planar Array on a Chip Using the MagArray Technology
Lateral Flow Immunoassay
Immuno-PCR Assay for Sensitive Detection of Proteins in Real Time
In Situ Proximity Ligation Assay (PLA)
Tyramide Signal Amplification for Immunofluorescent Enhancement
Correlative Microscopy for Localization of Proteins In Situ: Pre-embedding Immuno-Electron Microscopy Using FluoroNanogold, Gold Enhancement, and Low-Temperature Resin
Multiplex ELISA Using Oligonucleotide Tethered Antibodies
Gas Plasma Surface Chemistry for Biological Assays