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Multiplex Fluorescent Bead-Based Immunoassay for the Detection of Cytokines, Chemokines, and Growth Factors

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Proteomic Profiling

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2261))

Abstract

The comprehensive analysis of serum cytokine levels can be challenging due to low sample volumes and time consuming when using single-target methods like enzyme-linked immunosorbent assay (ELISA). Bead-based detection systems allow the simultaneous detection of multiple analytes using minimal sample volumes. Here we describe the use of a multiplex cytokine, chemokine, and growth factor assay for mouse cytokines in a 96-well format. This assay is based on antibody-coupled fluorescent magnetic beads combined with biotinylated secondary detection antibody followed by fluorescent-tagged streptavidin in a sandwich-like composition. Final assay readout provides the concentrations of 23 different cytokines, chemokines, and growth factors in up to 76 samples.

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Authors and Affiliations

  1. Institute of Comparative Molecular Endocrinology (CME), Ulm University, Ulm, Germany

    Jonathan M. Preuss, Ute Burret & Sabine Vettorazzi

Authors
  1. Jonathan M. Preuss
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  2. Ute Burret
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  3. Sabine Vettorazzi
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Corresponding authors

Correspondence to Jonathan M. Preuss or Sabine Vettorazzi .

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Editors and Affiliations

  1. Bio-Rad Laboratories GmbH, Feldkirchen, Germany

    Anton Posch

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Preuss, J.M., Burret, U., Vettorazzi, S. (2021). Multiplex Fluorescent Bead-Based Immunoassay for the Detection of Cytokines, Chemokines, and Growth Factors. In: Posch, A. (eds) Proteomic Profiling. Methods in Molecular Biology, vol 2261. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1186-9_15

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  • DOI: https://doi.org/10.1007/978-1-0716-1186-9_15

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1185-2

  • Online ISBN: 978-1-0716-1186-9

  • eBook Packages: Springer Protocols

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