Preparation of gold nanocluster bioconjugates for electron microscopy

Methods Mol Biol. 2013:950:293-311. doi: 10.1007/978-1-62703-137-0_17.

Abstract

In this chapter, we describe types of gold nanoparticle-biomolecule conjugates and their use in electron microscopy. Included are two detailed protocols for labeling an IgG antibody with gold monolayer protected clusters. The first approach is a direct bonding approach that utilizes the ligand place exchange reaction. The second approach describes NHS-EDC coupling of Au(144)(pMBA)(60) with IgG. Also included are various characterization techniques for determining labeling efficiency.

MeSH terms

  • Benzoates / chemistry
  • Carbodiimides / chemistry
  • Chemical Fractionation
  • Cryoelectron Microscopy
  • Electrophoresis, Polyacrylamide Gel
  • Gold / chemistry*
  • Immunoglobulin G / metabolism*
  • Ligands
  • Metal Nanoparticles / ultrastructure*
  • Microscopy, Electron / methods*
  • Sulfhydryl Compounds / chemistry

Substances

  • 1-ethyl-3-(3-(diethylamino)propyl)carbodiimide
  • Benzoates
  • Carbodiimides
  • Immunoglobulin G
  • Ligands
  • Sulfhydryl Compounds
  • 4-mercaptobenzoate
  • Gold