Flow Cytometry-Based Bead-Binding Assay for Measuring Receptor Ligand Specificity

Methods Mol Biol. 2016:1390:121-9. doi: 10.1007/978-1-4939-3335-8_8.

Abstract

In this chapter we describe a fluorescent bead-binding assay, which is an efficient and feasible method to measure interaction between ligands and receptors on cells. In principle, any ligand can be coated on fluorescent beads either directly or via antibodies. Binding between ligand-coated beads and cells can be measured by flow cytometry, which results in an easily quantifiable readout. Furthermore, it allows measuring of binding by specific cell subsets within a mixed cell population. Overall, this method is a convenient and easily standardized assay for measuring binding.

Keywords: Adhesion assay; Binding; C-type lectin receptors; CLRs; Ligand specificity; PRR.

MeSH terms

  • Animals
  • CHO Cells
  • Cricetulus
  • Flow Cytometry / methods*
  • Fluorescent Dyes
  • Ligands*
  • Microspheres
  • Protein Binding
  • Receptors, Cell Surface / metabolism*
  • Streptavidin

Substances

  • Fluorescent Dyes
  • Ligands
  • Receptors, Cell Surface
  • Streptavidin