Conjugation of Antibodies to Horseradish Peroxidase

Cold Spring Harb Protoc. 2019 Sep 3;2019(9). doi: 10.1101/pdb.prot099309.

Abstract

Antibodies conjugated with horseradish peroxidase (HRP) are one of the most widely used bioreagents in the biological sciences. This protocol is a basic method for adding HRP to a thiolated antibody and can be adapted for use with different cross-linkers. Conjugation methods usually focus on linking through the lysines on HRP because there are only six of them and their modification does not adversely affect enzyme activity.

MeSH terms

  • Antibodies / metabolism*
  • Horseradish Peroxidase / immunology*
  • Maleimides / metabolism
  • Succinimides / metabolism
  • Sulfides / metabolism

Substances

  • Antibodies
  • Maleimides
  • Succinimides
  • Sulfides
  • maleimide
  • N-hydroxysuccinimide S-acetylthioacetate
  • Horseradish Peroxidase