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Updated: Feb 25, 2026

A Simple Method to Identify Kinases That Regulate Embryonic Stem Cell Pluripotency by High-throughput Inhibitor Screening

A Simple Method to Identify Kinases That Regulate Embryonic Stem Cell Pluripotency by High-throughput Inhibitor Screening

Published on: May 12, 2017

A multiplexed screening method for pluripotency.

Alexander Plotnikov¹, Noga Kozer¹, Vladislav Krupalnik²

¹The Nancy & Stephen Grand Israel National Center for Personalized Medicine, Weizmann Institute of Science, Rehovot, Israel.

Stem Cell Research

|July 31, 2017

Summary

This summary is machine-generated.

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We developed a simple, inexpensive luminescence assay to measure intracellular Alkaline Phosphatase (ALP) levels and cell number simultaneously. This method is effective for analyzing somatic reprogramming and high-throughput screening.

Area of Science:

Biochemistry
Cell Biology
Stem Cell Research

Background:

Alkaline Phosphatase (ALP) level measurement is crucial in clinical diagnostics and biological research.
Existing methods for ALP quantification can be complex or costly.
Multiplexed and normalized intracellular ALP measurements are needed for advanced cellular analysis.

Purpose of the Study:

To present a simple, inexpensive, and multiplexed luminescence-based method for quantifying intracellular Alkaline Phosphatase (ALP) levels.
To enable simultaneous normalization of ALP levels with cell number in a single well.
To demonstrate the method's utility in analyzing somatic cell reprogramming.

Main Methods:

Utilized two commercially available reagents for sequential luminescence readouts.

Keywords:

ATP Alkaline Differentiation Drug Phosphatase Screening

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Last Updated: Feb 25, 2026

A Simple Method to Identify Kinases That Regulate Embryonic Stem Cell Pluripotency by High-throughput Inhibitor Screening

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Quantified intracellular ALP levels in one step.

Quantified cell number in a sequential step within the same sample well.

Main Results:

Successfully developed a luminescence-based assay for intracellular ALP quantification.
Demonstrated simultaneous normalization of ALP levels with cell number.
Applied the method to detect and analyze somatic reprogramming into pluripotent stem cells.

Conclusions:

The developed method offers a simple, cost-effective, and multiplexed approach for measuring intracellular ALP.
This assay is highly suitable for high-throughput screening (HTS) applications.
The method provides a valuable tool for stem cell research and reprogramming analysis.