Global Detection of Proteins by Label-Based Antibody Array

Methods Mol Biol. 2021:2237:45-53. doi: 10.1007/978-1-0716-1064-0_4.

Abstract

Because of narrow availability of antibody pairs and potential cross-reactivity between antibodies, the development of sandwich-based antibody arrays which need a pair of antibodies for each target has been restricted to higher density resulting in limited proteomic breadth of detection. Label-based array is one way to overcome this obstacle by directly labeling all targets in samples with fluorescent dyes such as Cy3 and Cy5. The labeled samples are then applied on the antibody array chip composed of capture antibodies. In this chapter, we will introduce this technology including array production and sample detection assay.

Keywords: Antibody array; Biotin labeling.

MeSH terms

  • Animals
  • Biotinylation / methods
  • Fluorescent Antibody Technique / methods*
  • Fluorescent Dyes / chemistry
  • Humans
  • Immunoassay / methods
  • Protein Array Analysis / methods*
  • Proteome / chemistry
  • Proteome / immunology
  • Proteomics / methods*

Substances

  • Fluorescent Dyes
  • Proteome