ROS/Superoxide Detection Assay Kit (Cell-based) (ab139476) (2024)

Overview

• Product name

  ROS/Superoxide Detection Assay Kit (Cell-based)
  See all Oxidative Stress kits

• Sample type

  Adherent cells, Suspension cells

• Assay type

  Cell-based

• Assay time

  0h 90m

• Product overview

  ROS/Superoxide Detection Assay Kit ab139476 is designed to directly monitor real time reactive oxygen species (ROS) production in live cells using fluorescence microscopy, flow cytometry, or microplate reader.

  
  

  The ROS/superoxide assayprotocol is based ontwo fluorescent dyes: Oxidative Stress Detection Reagent (Green, Ex/Em 490/525 nm) for the detection of total ROS, and Superoxide Detection Reagent (Orange, Ex/Em 550/620 nm).

  
  

  Through the combination of these two specific fluorescent probes, the kit provides a simple and specific assay for the real-time measurement of global levels of total reactive oxygen species (ROS), and of superoxides, in living cells. The kit is compatible with the major components of tissue culture media (phenol red, FBS and BSA).

  
  

  ROS/superoxide assay protocol summary:
  - add ROS inhibitor to negative controlcells and incubate for 30 min
  - addROS/superoxide detection mix (with the addition of ROS inducer for positive control cells) and incubate for 30-60 min
  - remove and discard the detection mix liquid
  - wash samples andanalyze with fluorescence microscope, or analyze by flow cytometry or microplate readerwithout washing

• Notes

  Reagents provided in the kit are sufficient for at least 200 tests using Fluorescent Microscopy or 50 tests using Flow Cytometry.

  Related products

  Review theoxidative stress marker and assay guide, or the fullmetabolism assay guideto learn about more assays for metabolites, metabolic enzymes, mitochondrial function, and oxidative stress, and also how to assay metabolic function in live cells using your plate reader.

  To measure reactive oxygen species within cells, we recommendDCFDA / H2DCFDA- Cellular ROS Assay Kit ab113851.Alternative ROS assays are available in orange (ab186028), red (ab186027), and deep red (ab186029). ab238535 is used to measure ROS in biofluids, culture supernatants and cell lysates.

  For assays designed to differentiate ROS, superoxides, and reactive nitrogen species: to assay ROS and superoxides use ab139476; to assay ROS, superoxides, and reactive nitrogen species use ab139473; to assay superoxides use ab219943.

• Platform

  Flow cytometer, Fluorescence microscope

Properties

• Storage instructions

  Please refer to protocols.

• Components	1 kit
  Oxidative Stress Detection Reagent (Green)	1 x 300nmole
  ROS Inducer (Pyocyanin 1 µmole)	1 vial
  ROS Inhibitor (N-acetyl-L-cysteine)	2 x 10mg
  Superoxide Detection Reagent (Orange)	1 x 300nmole
  Wash Buffer Salts	1 pack

• Research areas

  • Kits/ Lysates/ Other
  • Kits
  • Cell Damage Kits
  • Oxidative stress

  • Metabolism
  • Pathways and Processes
  • Redox metabolism
  • Oxidative stress

  • Kits/ Lysates/ Other
  • Kits
  • Cell Metabolism Kits
  • Oxidative Stress Assay Kits
  • Oxidative Stress

• Functional Studies - ROS/Superoxide Detection Assay Kit (Cell-based) (ab139476)M.Buchholz et al., BMC Cancer., Fig 5, doi:10.1186

  The level of ROS was analyzed in untreated compared to 2250 treated cells, additional NAC treatment served as a neg. Control

• Fluorescence Microscopy Profiling

  Profiling of reactive oxygen species formation by Fluorescence Microscopy was achieved in HeLa cells loaded with ROS/Superoxide detection reagents (ab139476) and treated with pyocyanin. General oxidative stress levels were monitored in the green channel, while superoxide production was detected in the orange channel. Pretreatment with NAC, a general ROS inhibitor, prevents formation of ROS.

• Flow Cytometry Analysis

  Jurkat cells were induced with 100 µM pyocyanin (general ROS inducer,panel B), 200 µM antimycin A (superoxide inducer, panel C) or 1 µM of t-butylhydroperoxide(peroxide inducer, panel D), stained with two color ROS Detection Kitand analyzed using flow cytometry. Untreated cells (panel A) were used as acontrol. Cell debris were ungated and compensation was performed using singlestained pyocyanin-treated samples. Red numbers reflect the percentage of the cellsin each quadrant.

• Protocol Booklet

Click here to view the general protocols

• SDS download

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• Datasheet download

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ab139476 has been referenced in 69 publications.

• Kanakkanthara A et al. Repurposing Ceritinib Induces DNA Damage and Enhances PARP Inhibitor Responses in High-Grade Serous Ovarian Carcinoma. Cancer Res 82:307-319 (2022). PubMed: 34810199
• Wu CH et al. 2-PMAP Ameliorates Cerebral Vasospasm and Brain Injury after Subarachnoid Hemorrhage by Regulating Neuro-Inflammation in Rats. Cells 11:N/A (2022). PubMed: 35053358
• Chen YL et al. Omega-3 fatty acids impair miR-1-3p-dependent Notch3 down-regulation and alleviate sepsis-induced intestinal injury. Mol Med 28:9 (2022). PubMed: 35090386
• Cui W et al. Inhibition of Autophagy Facilitates XY03-EA-Mediated Neuroprotection against the Cerebral Ischemia/Reperfusion Injury in Rats. Oxid Med Cell Longev 2022:7013299 (2022). PubMed: 35401933
• Liu B & Wang H Oxaliplatin induces ferroptosis and oxidative stress in HT29 colorectal cancer cells by inhibiting the Nrf2 signaling pathway. Exp Ther Med 23:394 (2022). PubMed: 35495610

View all Publications for this product