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ONE-Glo™ + Tox Luciferase Reporter and Cell Viability Assay

E7110

Sensitive Detection of Reporter Activity and Cell Health

  • Measures cell viability and firefly luciferase activity in the same assay well
  • Easy to use—simply add reagent, mix and read
  • Scalable to meet throughput needs, up to 1,536-well format

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Catalog number selected: E7110

$ 231.00 Your price: Log In

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ONE-Glo™ + Tox Luciferase Reporter and Cell Viability Assay
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$ 231.00
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Understand Reporter Gene Activity in the Context of Cell Viability

The ONE-Glo™ + Tox Assay combines luciferase assay chemistry with a cell viability marker to better understand reporter gene expression in the context of cell health. The assay uses a two-step, addition-only process to make these measurements in a single well of a plate, negating the need to run parallel assays. The simple "add-mix-read" assay format and optimized reagents combine to create a flexible, automation-friendly assay that can be scaled to meet your needs.

Step 1: Cell Viability Assay

In the first part of the assay, a nonlytic fluorescence-based method (CellTiter-Fluor™ Cell Viability Assay) is used to measure the relative number of live cells in culture. The CellTiter-Fluor™ Assay measures a conserved and constitutive protease activity within living cells. This live-cell protease activity is restricted to intact viable cells and is measured using a fluorogenic, cell-permeant peptide substrate (glycylphenylalanyl-aminofluorocoumarin; GF-AFC). The substrate enters intact cells where it is cleaved by the live-cell protease to generate a fluorescent signal proportional to the number of living cells. The live-cell protease becomes inactive upon loss of cell membrane integrity and leakage into the surrounding culture medium. Fluorescence of the free AFC fluorophore is measured with a microplate reader or CCD imager using an excitation wavelength of 380–400nm and emission wavelength of 505nm.

Step 2: ONE-Glo™ Luciferase Assay

The second part of the assay uses the ONE-Glo™ Luciferase Assay System to quantify firefly luciferase gene expression from cells expressing the reporter. Ideally suited for high- and ultrahigh-throughput applications, the ONE-Glo™ Assay Reagent contains a fluoroluciferin substrate that gives increased stability and greater tolerance of sample components, and has less odor than standard luciferase assay reagents. Luminescence is measured with a microplate reader or CCD imager.

Reference

Niles, A.L. et al. (2007) A homogeneous assay to measure live and dead cells in the same sample by detecting different protease markers. Anal. Biochem. 366, 197–206.

10471MA.eps
Schematic of the ONE-Glo™+Tox Luciferase Reporter and Cell Viability Assay.

Example ONE-Glo™ + Tox Assay Data

Ionomycin Titration in 96- and 384-well Plates

In the experiment shown here, 1 × 104 (96-well plate; Panel A) or 5 × 103 (384-well plate; Panel B) cells expressing NFAT response element were treated with serial titrations of ionomycin in the presence of PMA for 6 hours. At specific concentrations, ionomycin and PMA work cooperatively to stimulate NFAT-dependent gene expression. However, higher concentrations of ionomycin result in cytotoxicity seen as a decrease in viability (fluorescence). A decrease in reporter expression (luciferase) activity is also observed due to the increase in cytotoxicity.


Ionomycin titration in standard 96- and 384-well plate formats.

Protocols

Specifications

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What's in the box?

Item Part # Size

ONE-Glo™ Luciferase Assay Buffer

E605A 1 x 10ml

ONE-Glo™ Luciferase Assay Substrate

E606A 1 x 1 vial

GF-AFC Substrate

G608A 1 x 10μl

Assay Buffer

G610A 1 x 10ml

SDS

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Certificate of Analysis

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Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

Storage Conditions

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Patents and Disclaimers

U.S. Pat. Nos. 7,416,854, 7,553,632 and other patents pending.

Patent Pending.

Certain applications of this product may require licenses from others.

Specifications

You are viewing: 10 plates Change Configuration

What's in the box?

Item Part # Size

ONE-Glo™ Luciferase Assay Buffer

E605B 1 x 100ml

ONE-Glo™ Luciferase Assay Substrate

E606B 1 x 1 vial

GF-AFC Substrate

G608B 2 x 50μl

Assay Buffer

G610B 1 x 50ml

SDS

Choose language:

Certificate of Analysis

Search for Specific Certificate:

View more results
No results
Loading…

Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

Storage Conditions

BB

Patents and Disclaimers

U.S. Pat. Nos. 7,416,854, 7,553,632 and other patents pending.

Patent Pending.

Certain applications of this product may require licenses from others.

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