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Updated: Jan 4, 2026

A Multiplexed Luciferase-based Screening Platform for Interrogating Cancer-associated Signal Transduction in Cultured Cells

A Multiplexed Luciferase-based Screening Platform for Interrogating Cancer-associated Signal Transduction in Cultured Cells

Published on: July 3, 2013

Multiplexing fluorogenic esterase-based viability assay with luciferase assays.

Kenji Ohgane¹, Hiromasa Yoshioka¹, Yuichi Hashimoto¹

¹Institute for Quantitative Biosciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-0032, Japan.

|November 1, 2019

Summary

This summary is machine-generated.

This multiplex assay enables sequential measurement of cell viability and luciferase activity in drug discovery screening. It streamlines workflows by eliminating the need for separate assays, reducing false positives and improving efficiency.

Keywords:

CytoRed CytoRed-luciferase multiplex assay Esterase Fluorogenic substrate Luciferase Multiplex assay Viability

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Last Updated: Jan 4, 2026

A Multiplexed Luciferase-based Screening Platform for Interrogating Cancer-associated Signal Transduction in Cultured Cells

A Multiplexed Luciferase-based Screening Platform for Interrogating Cancer-associated Signal Transduction in Cultured Cells

Published on: July 3, 2013

High-throughput Functional Screening using a Homemade Dual-glow Luciferase Assay

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Published on: June 1, 2014

Studying Membrane Biogenesis with a Luciferase-Based Reporter Gene Assay

Studying Membrane Biogenesis with a Luciferase-Based Reporter Gene Assay

Published on: September 7, 2008

Area of Science:

Biochemistry
Cell Biology
Drug Discovery

Background:

Luciferase reporter assays are standard in cell-based drug discovery.
Assessing compound cytotoxicity alongside reporter activity is crucial for accurate results.
Existing methods often require separate assays or parallel sample handling.

Purpose of the Study:

To develop a multiplex assay for sequential measurement of cell viability and luciferase activity.
To streamline drug screening by integrating cytotoxicity assessment.
To provide a protocol suitable for cells with stable luciferase expression.

Main Methods:

Utilized a fluorogenic esterase substrate (CytoRed) for cell viability assessment.
Developed a sequential measurement protocol within a multi-well-plate format.
Applied the assay to cells stably expressing luciferase constructs.

Main Results:

Successfully demonstrated sequential measurement of esterase activity (cell viability) followed by luciferase activity in the same sample.
Eliminated the requirement for parallel viability or protein assays using separate sample aliquots.
Validated the protocol's utility for cell lines with stable luciferase expression.

Conclusions:

The developed multiplex assay offers an efficient method for simultaneous assessment of cell viability and luciferase activity.
This protocol enhances drug discovery screening by reducing false positives and simplifying experimental procedures.
It is particularly advantageous for applications involving stably transfected cell lines.