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Related Experiment Video

Updated: May 9, 2026

Identification of Kinase-substrate Pairs Using High Throughput Screening
11:13

Identification of Kinase-substrate Pairs Using High Throughput Screening

Published on: August 29, 2015

Phosphatase high-throughput screening assay design and selection.

Eduard A Sergienko1

  • 1Conrad Prebys Center for Chemical Genomics, Sanford-Burnham Medical Research Institute, La Jolla, CA, USA.

Methods in Molecular Biology (Clifton, N.J.)
|July 18, 2013
PubMed
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Phosphatase assays measure enzyme activity using various biochemical methods. Developing comprehensive assay panels is key to discovering effective and specific phosphatase modulators for biological applications.

Area of Science:

  • Biochemistry
  • Enzymology

Background:

  • Phosphatases are enzymes that remove phosphate groups from diverse substrates.
  • Existing biochemical assays for measuring phosphatase activity include spectrophotometric methods and inorganic phosphate quantitation.
  • Current assays often struggle to accurately reflect native reactions or support high-throughput screening.

Purpose of the Study:

  • To review existing biochemical approaches for measuring phosphatase activity.
  • To highlight the limitations of current assay types in reflecting native reactions and enabling high-throughput screening.
  • To propose the utilization of comprehensive assay panels and novel screening approaches for identifying biologically relevant phosphatase modulators.

Main Methods:

  • Review of spectrophotometric methods using chromogenic, fluorogenic, and luminogenic substrates.

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Last Updated: May 9, 2026

Identification of Kinase-substrate Pairs Using High Throughput Screening
11:13

Identification of Kinase-substrate Pairs Using High Throughput Screening

Published on: August 29, 2015

Multi-enzyme Screening Using a High-throughput Genetic Enzyme Screening System
08:10

Multi-enzyme Screening Using a High-throughput Genetic Enzyme Screening System

Published on: August 8, 2016

  • Analysis of phosphate detection methods for quantitating inorganic phosphate.
  • Discussion of cell-based model systems and novel screening approaches.

    • Main Results:

    • Spectrophotometric and inorganic phosphate assays are broadly used but have limitations.
    • Artificial substrates may not accurately mimic native enzyme kinetics and substrate interactions.
    • Current methods often fail to provide biologically relevant phosphatase modulators or support high-throughput screening.

    Conclusions:

    • Comprehensive phosphatase assay panels combining biochemical assays and cell-based systems are crucial.
    • Novel screening approaches are needed to identify potent, selective, and biologically active phosphatase modulators.
    • Addressing the limitations of current assays will advance the discovery of novel phosphatase-targeted therapeutics.