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Related Experiment Video

Updated: Jun 8, 2026

Probing High-density Functional Protein Microarrays to Detect Protein-protein Interactions
08:07

Probing High-density Functional Protein Microarrays to Detect Protein-protein Interactions

Published on: August 2, 2015

A peptide microarray for detecting protein kinase activity in cell lysates.

Xiaoming Han1, Yoshiki Katayama

  • 1Graduate School of System Life Sciences, Kyushu University, Fukuoka, Japan.

Methods in Molecular Biology (Clifton, N.J.)
|September 22, 2010
PubMed
Summary

We developed a sensitive peptide microarray to detect intracellular protein kinase (PK) activity. This high-throughput assay is valuable for disease diagnosis and drug discovery.

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Assay Development

Background:

  • Protein kinases (PKs) are crucial targets in disease diagnosis and drug discovery.
  • Detecting intracellular PK activity is essential for understanding cellular processes and disease mechanisms.

Purpose of the Study:

  • To develop a sensitive peptide microarray for detecting intracellular PK activity.
  • To establish a high-throughput method for kinase assays with potential applications in disease diagnosis and drug discovery.

Main Methods:

  • Immobilizing substrate peptides on glutaraldehyde-premodified glass slides using an automated microarray spotter.
  • Detecting phosphorylated peptides using fluorescence-labeled antiphosphotyrosine antibodies (for tyrosine kinases) or Phos-tag biotin and fluorescence-labeled streptavidin (for serine/threonine kinases).

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Last Updated: Jun 8, 2026

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  • Scanning fluorescence signals with an automatic microarray scanner.

    • Main Results:

    • The developed peptide microarray system demonstrates simple peptide immobilization and requires low sample volumes.
    • The system provides a high-density array with high sensitivity for detecting PK activities in cell lysates.
    • The assay effectively distinguishes between tyrosine and serine/threonine kinase activities.

    Conclusions:

    • The peptide microarray system offers a sensitive and high-throughput method for assessing intracellular PK activity.
    • This technology has significant potential for applications in disease diagnosis and accelerating drug discovery efforts.
    • The assay's simplicity, low sample requirement, and high sensitivity make it a valuable tool for kinase research.