ROS-Glo™ H2O2 Assay

Part Numbers: G8820, G8821

G8820_ROS-Glo-H2O2-Assay-10ml
automation-support-promo
customize-this-small

過酸化水素 (H2O2) を特異的、直接的に検出する発光アッセイ

  • 2種類の試薬を順次添加するだけのシンプルなプロトコール
  • H2O2 と直接反応する基質を使用するためカップリング酵素 HRPも不要

Size

選択製品のカタログ番号: G8820

Please Enquire
This product is discontinued
Request a Sample
Not available in all countries
This product is available under our Early Access program - Learn More
This product is available under our Catalog (FT) program - Learn More
ROS-Glo™ H2O2 Assay
10ml
Your price:
Change Configuration

ROS-Glo™ H2O2 Assayは活性酸素種(ROS)である過酸化水素(H2O2)レベルを測定する迅速で高感度なホモジニアス生物発光アッセイで、培養細胞あるいは特定の酵素反応より直接測定することができます。ルシフェリン誘導体基質をサンプルとともにインキュベートするとH2O2と直接反応してルシフェリン前駆体が生成されます。ROS-Glo™ Detection Solutionの添加によりこの前駆体がルシフェリンに変換され、 試薬に含まれるUltra-Glo™ Recombinant Luciferaseがサンプル中に存在するH2O2レベルに比例した発光シグナルを生じます。


  • 直接的な細胞ベースの検出:血清の有無にかかわらず様々な培地で使用でき、アッセイ前に培養細胞から培地を除去する必要もありません。
  • シンプルで迅速なアッセイ:2種類の試薬を加えるだけのシンプルなホモジニアスアッセイによりサンプルの前処理も不要。アッセイは試薬添加後2時間以内に完了。
  • HRP未使用:ROS-Glo™ H2O2 SubstrateはH2O2と直接反応し、カップリング酵素としてホースラディッシュペルオキシダーゼ(HRP)を使用しないため、HRP阻害剤による偽陽性を排除。
  • 自動化に適応:96~384ウェルプレート形式へ容易に対応。
  • 柔軟なアッセイ:細胞ベースあるいは酵素ベースの両方で化合物スクリーニングに使用可能。
  • マルチプレックスに対応:リアルタイム細胞毒性アッセイCellTox™ Green Cytotoxicity Assayと併用した同一ウェルでのマルチプレックスアッセイあるいは生存性アッセイとのマルチアッセイでウエルあたりのデータをより多く入手でき、細胞培養のコストも低減。

 

  • 培養細胞サンプルの過酸化水素レベル変化の直接的な測定
  • H2O2を生成あるいは消費する酵素活性の測定
  • 培養細胞内あるいは酵素反応におけるROSレベルに変化を与える低分子阻害剤/誘導剤の同定

※10mlは96ウェルプレートの場合100ウェル分(100µl/ウェル使用)、384ウェルプレートの場合400ウェル分(25µl/ウェル使用)に相当

【毒劇物】対象品 ※SDSは本ページ下部より入手ください。

ROS-Glo™ H₂O₂ Assay Chemistry

11359MA-W

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

11360MA-W

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

11901MA-W

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

11902MA-W

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 

11586MA-W

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.

Case Study

Automated Assay Workflow for Mitochondrial Dysfunction

Challenge:
Louise Young, a research fellow at the University of Strathclyde, studies mitochondria’s role in Alzheimer’s and cancer. Her team needed a reliable, reproducible method to measure ROS output while screening compounds to decrease ROS levels. Traditional assays had wash steps, causing variability.

Result:
Using ROS-Glo™ H2O2 Assay, the team achieved faster turnaround with a no-wash protocol, greater reproducibility through luminescent readout and improved efficiency for quicker data collection.

Their workflow also includes RealTime-Glo™ Annexin V Assay for apoptosis/necrosis and CellTiter-Glo® 2.0 Assay for cell viability. Several compounds successfully reduced ROS, guiding future research into their protein targets.

Download the Case Study

製品仕様

You are viewing: G8820 Change Configuration

選択製品の構成品内容

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

分析証明書

Search by lot number

使用制限

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

保存条件

BB

製品仕様

You are viewing: G8821 Change Configuration

選択製品の構成品内容

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

分析証明書

Search by lot number

使用制限

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

保存条件

BB

Let's find the product that meets your needs.

Talk to a Scientist

scientist-uk-manos

Manos

UK