Protein Arrays pp 161-172

Part of the Methods in Molecular Biology book series (MIMB, volume 264)

A Protein Microarray ELISA for Screening Biological Fluids

  • Susan M. Varnum
  • Ronald L. Woodbury
  • Richard C. Zangar

Abstract

Protein microarrays permit the simultaneous measurement of many proteins in a small sample volume and therefore provide an attractive approach for the quantitative measurement of proteins in biological fluids, including serum. This chapter describes a microarray enzyme-linked immunosorbent assay (ELISA). Capture antibodies are immobilized onto a glass surface; the covalently attached antibodies bind a specific antigen from a sample overlaying the array. A second, biotinylated antibody that recognizes the same antigen as the first antibody, but at a different epitope, is then used for detection. Detection is based on an enzymatic signal-enhancement method known as tyramide signal amplification (TSA). By coupling a microarray-ELISA format with the signal amplification of tyramide deposition, the assay sensitivity is as low as sub-pg/mL.

Key Words

Microarray ELISA tyramide antibody biological fluids serum 

Copyright information

© Humana Press Inc., Totowa, NJ 2004

Authors and Affiliations

  • Susan M. Varnum
    • 1
  • Ronald L. Woodbury
    • 2
  • Richard C. Zangar
    • 1
  1. 1.Biological Sciences DivisionPacific Northwest National LaboratoryRichland
  2. 2.Biological Sciences DivisionPacific Northwest National LaboratoryBattelle, Richland

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