Protocol

Protein Arrays: Labeling the Protein and Probing the Array for Protein-Protein Interactions

This protocol was adapted from "Characterization of Protein Complexes," Chapter 10, in Proteins and Proteomics (ed. Simpson). Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA, 2003.

INTRODUCTION

To detect protein-protein interactions, a protein microarray can be incubated with a fluorescently labeled protein. After the array is washed, stable interactions can be identified by scanning the slide for fluorescent spots. This protocol uses cyanine-3 (Cy3) or cyanine-5 (Cy5) monofunctionally activated dyes to label the probe proteins.