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ROS-Glo™ H2O2 Assay

Specific, Direct Detection of Hydrogen Peroxide (H2O2)

  • Fast assay involves a simple two-reagent-addition protocol
  • Substrate reacts directly with H2O2; no need for HRP as a coupling enzyme

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Catalog number selected: G8820

$ 377.00
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ROS-Glo™ H2O2 Assay
10ml
$ 377.00
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The ROS-Glo™ H₂O₂ Assay is a homogeneous, fast and sensitive bioluminescent assay that measures the level of hydrogen peroxide (H₂O₂), a reactive oxygen species (ROS), directly in cell culture or in defined enzyme reactions. A derivatized luciferin substrate is incubated with sample and reacts directly with H₂O₂ to generate a luciferin precursor. Addition of ROS-Glo™ Detection Solution converts the precursor to luciferin and provides Ultra-Glo™ Recombinant Luciferase to produce light signal that is proportional to the level of H₂O₂ present in the sample.

ROS-Glo™ H₂O₂ Assay chemistry

The H₂O₂ Substrate reacts directly with H₂O₂ to create the Luciferin Precursor. The ROS-Glo™ Substrate Detection Solution converts the Luciferin Precursor to luciferin and also provides the Ultra-Glo™ Luciferase to produce light. Light signal is proportional to the amount of H₂O₂ in the sample well.

Overview of the ROS-Glo™ H₂O₂ Assay Protocol for Cell-based or Biochemical Detection

The assay follows a simple two-reagent-addition protocol that does not require sample manipulation. The ROS-Glo™ H₂O₂ Substrate can be added to assay wells with the treatment or at the end of the treatment incubation period, and data can be recorded in less than 2 hours after reagent addition.

Perform Enzymatic or Cell-based Assays in 96- and 384-well Plate Formats

ROS Induction in Cultured Cells

Menadione treatment of K562 cells resulted in a concentration-dependent ROS increase as detected with the ROS-Glo™ H₂O₂ Assay. The assay can be performed in various cell culture media with or without serum, eliminating the need to remove the media from cultured cells before performing the assay.

NADH Oxidase Enzymatic Activity

NADH Oxidase enzymatic activity was determined by incubating increasing concentrations of NADH oxidase with NADH in enzyme reaction buffer and measuring the H₂O₂ produced with the ROS-Glo™ H₂O₂ Assay. After appropriate levels of NADH oxidase and NADH substrate were determined, the assay was used to identify inhibitors of the enzyme in a chemical library by looking for decreased luminescence.

Multiplex With a Cytotoxicity Assay for More Informative Data Per Well 

HepG2 cells were treated with either ROS-generating compounds (menadione or pyrogallol) or a cytotoxicity inducing reagent (digitonin) and incubated at 37°C for 2 hours. CellTox™ Green Dye (membrane impermeable DNA binding dye as an indicator of cytotoxicity) and H₂O₂ Substrate were added at the time of dosing. After incubation fluorescence from the CellTox™ Green Dye was measured followed by addition of ROS-Glo™ Detection Solution. Luminescence signal from the ROS-Glo™ Assay was measured following 20‑minute incubation.

Cat.# G8820 contains sufficient reagents for 100 assays at 100µl/assay in 96-well plates or 400 assays at 25µl/assay in 384-well plate format. Cat.# G8821 contains sufficient reagents for 500 assays at 100µl/assay in 96-well plates or 2,000 assays at 25µl/assay in 384-well plate format.

Specifications

You are viewing: G8820 Change Configuration

What's in the box?

Item Part # Size

H2O2 Substrate, 10mM

G882A 1 × 40μl

Signal Enhancer Solution

G883A 1 × 100μl

H2O2 Substrate Dilution Buffer

G922A 1 × 2ml

d-Cysteine, 100X

V251A 1 × 100μl

Luciferin Detection Reagent

V859A 1 × 1 each

Reconstitution Buffer

V865A 1 × 10ml

SDS

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Download SDSPDF (264 KB) – English (United States)

Certificate of Analysis

See all certificates for G8820

Use Restrictions

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

Storage Conditions

BB

Patents and Disclaimers

European Pat. No. 1131441 and Japanese Pat. No. 4520084.

Patent Pending.

Specifications

You are viewing: G8821 Change Configuration

What's in the box?

Item Part # Size

H2O2 Substrate, 10mM

G882B 1 × 200μl

Signal Enhancer Solution

G883B 1 × 500μl

H2O2 Substrate Dilution Buffer

G922B 1 × 10ml

d-Cysteine, 100X

V251B 1 × 500μl

Luciferin Detection Reagent

V859B 1 × 1 each

Reconstitution Buffer

V865B 1 × 50ml

SDS

Choose language:

Download SDSPDF (240 KB) – English (United States)

Certificate of Analysis

See all certificates for G8821

Use Restrictions

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

Storage Conditions

BB

Patents and Disclaimers

European Pat. No. 1131441 and Japanese Pat. No. 4520084.

Patent Pending.

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