A capture coupling method for the covalent immobilization of hexahistidine tagged proteins for surface plasmon resonance

Methods Mol Biol. 2010:627:91-100. doi: 10.1007/978-1-60761-670-2_5.

Abstract

Surface plasmon resonance (SPR) is a robust method to detect and quantify macromolecular interactions; however, to measure binding interactions, one component must be immobilized on a sensor surface. This is typically achieved using covalent immobilization via free amines or thiols, or noncovalent immobilization using high-affinity interactions such as biotin/streptavidin or antibody/antigen. In this chapter we describe a robust method to covalently immobilize His(6) fusion proteins on the sensor surface for SPR analysis.

MeSH terms

  • Amines / chemistry
  • Buffers
  • GTP-Binding Proteins / analysis
  • GTP-Binding Proteins / chemistry
  • GTP-Binding Proteins / metabolism
  • Histidine / metabolism*
  • Immobilized Proteins / analysis*
  • Immobilized Proteins / chemistry*
  • Immobilized Proteins / metabolism
  • Oligopeptides / metabolism*
  • Protein Multimerization
  • Protein Structure, Quaternary
  • Reproducibility of Results
  • Surface Plasmon Resonance / methods*

Substances

  • Amines
  • Buffers
  • His-His-His-His-His-His
  • Immobilized Proteins
  • Oligopeptides
  • Histidine
  • GTP-Binding Proteins