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Related Experiment Video

Updated: Jun 2, 2026

An Optimized Single-Molecule Pull-Down Assay for Quantification of Protein Phosphorylation
07:45

An Optimized Single-Molecule Pull-Down Assay for Quantification of Protein Phosphorylation

Published on: June 6, 2022

Microarray-based kinetic colorimetric detection for quantitative multiplex protein phosphorylation analysis.

Pavlo Holenya1, Igor Kitanovic, Florian Heigwer

  • 1Department of Biology, Institut für Pharmazie und molekulare Biotechnologie, Ruperto-Carola University of Heidelberg, Heidelberg, Germany.

Proteomics
|April 19, 2011
PubMed
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This study introduces a kinetic colorimetric detection method for enzyme-linked immunosorbent assay (ELISA) microarrays, improving quantitative accuracy for phosphorylated proteins. The new approach extends the detection range significantly, enabling precise measurement of multiple targets simultaneously.

Area of Science:

  • Biochemistry
  • Analytical Chemistry
  • Molecular Biology

Background:

  • Colorimetric detection in protein microarrays with enzymatic amplification often yields non-linear signals, limiting quantitative accuracy.
  • Existing methods struggle with a broad dynamic range for analyte concentration, hindering precise measurements.

Purpose of the Study:

  • To develop a kinetic colorimetric detection protocol to extend the quantitative range of ELISA microarrays.
  • To enable accurate simultaneous measurement of multiple phosphorylated proteins.

Main Methods:

  • Developed a novel kinetic colorimetric detection protocol for ELISA microarrays.
  • Utilized ArrayTube™ and ArrayStrip™ platforms for analysis.
  • Calibrated microarrays over a four-orders-of-magnitude concentration range with a picomolar threshold.

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Published on: September 19, 2018

Kinase Inhibitor Screening In Self-assembled Human Protein Microarrays
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Kinase Inhibitor Screening In Self-assembled Human Protein Microarrays

Published on: October 23, 2019

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Last Updated: Jun 2, 2026

An Optimized Single-Molecule Pull-Down Assay for Quantification of Protein Phosphorylation
07:45

An Optimized Single-Molecule Pull-Down Assay for Quantification of Protein Phosphorylation

Published on: June 6, 2022

Assessment of Resistance to Tyrosine Kinase Inhibitors by an Interrogation of Signal Transduction Pathways by Antibody Arrays
07:42

Assessment of Resistance to Tyrosine Kinase Inhibitors by an Interrogation of Signal Transduction Pathways by Antibody Arrays

Published on: September 19, 2018

Kinase Inhibitor Screening In Self-assembled Human Protein Microarrays
13:22

Kinase Inhibitor Screening In Self-assembled Human Protein Microarrays

Published on: October 23, 2019

Main Results:

  • Achieved a significantly extended detection range for microarrays.
  • Demonstrated accurate quantification over four orders of magnitude of analyte concentration.
  • Successfully performed simultaneous quantitative measurement of 15 phosphorylated proteins on a single chip.

Conclusions:

  • The kinetic colorimetric detection protocol overcomes limitations of traditional methods for protein microarrays.
  • This approach enhances the accuracy and range of quantitative readouts for phosphorylated protein analysis.
  • Enables high-throughput, multiplexed quantification of biomarkers.