Handbook of ELISPOT

1. Front Matter

   Pages i-xiv

   PDF

2. Elispot Assay Setup and Data Analysis

   1. Front Matter

      Pages 1-1

      PDF

   2. ELISPOT Assay

      • Jonathon D. Sedgwick

      Pages 3-14

   3. Chemistry and Biology of the ELISPOT Assay

      • Alexander E. Kalyuzhny

      Pages 15-31

   4. Membranes and Membrane Plates Used in ELISPOT

      • Alan J. Weiss

      Pages 33-49

   5. Standardization and Validation Issues of the ELISPOT Assay

      • Sylvia Janetzki, Josephine H. Cox, Neal Oden, Guido Ferrari

      Pages 51-86

   6. A Cell-Detachment Solution Can Reduce Background Staining in the ELISPOT Assay

      • Angela Grant, Sarah Palzer, Chris Hartnett, Tanya Bailey, Monica Tsang, Alexander E. Kalyuzhny

      Pages 87-94

   7. Isolation of Subsets of Immune Cells

      • Carrie E. Peters, Steven M. Woodside, Allen C. Eaves

      Pages 95-115

   8. Image Analysis and Data Management of ELISPOT Assay Results

      • Paul Viktor Lehmann

      Pages 117-131

   9. High Resolution as a Key Feature to Perform Accurate ELISPOT Measurements Using Zeiss KS ELISPOT Readers

      • Wolf Malkusch

      Pages 133-150

3. Elispot Assay in Animal Models

   1. Front Matter

      Pages 151-151

      PDF

   2. Improving the Sensitivity of the ELISPOT Analyses of Antigen-Specific Cellular Immune Responses in Rhesus Macaques

      • K. Jagannadha Sastry, Pramod N. Nehete, Bharti Nehete

      Pages 153-165

   3. Feline Cytokine ELISPOT

      • Sushila K. Nordone, Rosemary Stevens, Alora S. LaVoy, Gregg A. Dean

      Pages 167-177

   4. Measuring T-Cell Function in Animal Models of Tuberculosis by ELISPOT

      • Vanja Lazarevic, Santosh Pawar, JoAnne Flynn

      Pages 179-189

   5. Interferon-γ ELISPOT Assay for the Quantitative Measurement of Antigen-Specific Murine CD8+ T-Cells

      • Geoffrey A. Cole

      Pages 191-203

4. Elispot Assays in Human Studies

   1. Front Matter

      Pages 205-205

      PDF

   2. Detection of Measles Virus-Specific Interferon-γ-Secreting T-Cells by ELISPOT

      • Jenna E. Ryan, Inna G. Ovsyannikova, Gregory A. Poland

      Pages 207-217

   3. Epitope Mapping in Multiple Sclerosis Using the ELISPOT Assay

      • Clara M. Pelfrey, Ioana R. Moldovan

      Pages 219-235

   4. Use of Interferon-γ ELISPOT in Monitoring Immune Responses in Humans

      • Mark Matijevic, Robert G. Urban

      Pages 237-251

   5. ELISPOT Determination of Interferon-γ T-Cell Frequencies in Patients With Autoimmune Sensorineural Hearing Loss

      • C. Arturo Solares, Vincent K. Tuohy

      Pages 253-260

In this first book dedicated entirely to the ELISPOT, a critical enzyme-linked immunospot assay used widely in biomedical research, recognized experts with first-hand experience detail how to design, perform, and analyze these assays. The readily reproducible techniques they provide cover a wide variety of topics, including the use of membrane-backed plates, the standardization and validation procedures, the removal of cells from ELISPOT plates, cell separation techniques, and the quantification of ELISPOT data. There are also numerous ELISPOT applications involving animal models, human cells, measles, multiple sclerosis, immune responses, multicytokine detection systems, and immunocytochemistry. Highlights include dual-color and multiplex ELISPOT assays, use of the ELISPOT assay on feline lymphocytes, standardization of the ELISPOT procedure, and combining the ELISPOT assay with immunohistochemistry.

• Antigen
• Termination
• antibody
• biology
• cell
• chemistry
• cytokine
• cytokines
• development
• immune response
• interferon
• lymphocytes
• membrane
• reproducible techniques
• tuberculosis

From the reviews:

"The book is divided into four sections, with a total of 20 chapters featuring step-by-step protocols that will enable both novice and experienced researchers to perform ELISPOT. Each chapter begins with a general Introduction, followed by a detailed Materials section and an easy-to-follow Methods section … . All the chapters are equally interesting and well presented … . a must for a cytochemist. This book will serve as a convenient reference manual for beginners and for experienced ELISPOT users." (Carlo Alberto Redi, European Journal of Histochemistry, December, 2005)

• R&D Systems Inc.,, Minneapolis

  Alexander E. Kalyuzhny

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