Protein Arrays: Probing the Array and Detecting Radioactive Spots for Kinase-Substrate 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
Protein arrays can be probed for potential substrates of protein kinases. To identify substrates of a kinase of interest, a protein microarray is incubated with that kinase and [γ-33P]ATP. Following an appropriate incubation, the array is washed, coated with a photographic emulsion, and incubated further; the emulsion is developed and then imaged with a scanning light microscope.
