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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.

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Images

Functional Studies - ROS/Superoxide Detection Assay Kit (Cell-based) (AB139476), expandable thumbnail
  • Flow Cytometry - ROS/Superoxide Detection Assay Kit (Cell-based) (AB139476), expandable thumbnail
  • Fluorescence Microscopy - ROS/Superoxide Detection Assay Kit (Cell-based) (AB139476), expandable thumbnail

Publications

Key facts

Sample types

Suspension cells, Adherent cells

Assay type

Cell-based

Assay time

90m

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What's included?

1 Kit
Components
Oxidative Stress Detection Reagent (Green)
1 x 300 nmol
ROS Inducer (Pyocyanin 1 µmole)
1 x 1 Vial
ROS Inhibitor (N-acetyl-L-cysteine)
2 x 10 mg
Superoxide Detection Reagent (Orange)
1 x 300 nmol
Wash Buffer Salts
1 x 1 Pack

Recommended products

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.

Key facts

Sample types

Suspension cells, Adherent cells

Assay type

Cell-based

Assay time

90m

Assay Platform

Flow cytometer, Fluorescence microscope

Storage

Shipped at conditions

Dry Ice

Appropriate short-term storage conditions

-80°C

Appropriate long-term storage conditions

-80°C

Storage information

-80°C

Notes

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 assay protocol is based on two 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 control cells and incubate for 30 min
- add ROS/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 and analyze with fluorescence microscope, or analyze by flow cytometry or microplate reader without washing

Reagents provided in the kit are sufficient for at least 200 tests using Fluorescent Microscopy or 50 tests using Flow Cytometry. **Related products** Review the , or the full to 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 recommend . Alternative ROS assays are available in orange (Cellular ROS Assay Kit (Orange) ab186028), red (Cellular ROS Assay Kit (Red) ab186027), and deep red (Cellular ROS Assay Kit (Deep Red) ab186029). DCF ROS/RNS Assay Kit (biofluids, culture supernatant, cell lysates) 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 Cellular ROS/RNS Assay Kit ab139473; to assay superoxides use Mitochondrial Superoxide Assay Kit (Fluorometric) ab219943.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

3 product images

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

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

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

  • Flow Cytometry - ROS/Superoxide Detection Assay Kit (Cell-based) (ab139476), expandable thumbnail

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

    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 Kit
    and analyzed using flow cytometry. Untreated cells (panel A) were used as a
    control. Cell debris were ungated and compensation was performed using single
    stained pyocyanin-treated samples. Red numbers reflect the percentage of the cells
    in each quadrant.

  • Fluorescence Microscopy - ROS/Superoxide Detection Assay Kit (Cell-based) (ab139476), expandable thumbnail

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

    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.

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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