ELISA pp 107-118 | Cite as

Multiplexed Microsphere Suspension Array-Based Immunoassays

Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1318)

Abstract

ELISA is an extremely powerful tool to detect analytes because of its sensitivity, selectivity, reproducibility and ease of use. Here we describe sandwich immunoassays performed in suspension on spectrally unique microspheres developed by Luminex. Luminex assays offer the benefit of multiplex analysis of large numbers of analytes in a single reaction. Because the microspheres are spectrally unique, many microspheres, each attached to various antibodies, can be added to a single sample. Luminex instruments can distinguish each microsphere and detect the intensity of a reporter signal for each microsphere. Results are reported in Median Fluorescent Intensities for each analyte. Luminex assays can be used to detect up to 500 analytes in a high-throughput format. Luminex refers to this technology as xMAP®. Here we describe a routine protocol for a Luminex immunoassay. Other Luminex assays would have to be optimized for specific conditions according to their use.

Key words

Microsphere Bead-based assay ELISA Immunoassay Antibody Multiplex 
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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Andrew Lin
    • 1
  • Alexandra Salvador
    • 2
  • J. Mark Carter
    • 2
  1. 1.United States Food and Drug Administration (FDA)AlamedaUSA
  2. 2.Agricultural Research ServicesU.S. Department of AgricultureAlbanyUSA

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