Cellular ROS/RNS Assay Kit (ab139473)

Reviews (1)Q&A (2)References (2)

Overview

  • Product name

    Cellular ROS/RNS Assay Kit
    See all Oxidative Stress kits

  • Sample type

    Adherent cells, Suspension cells

  • Assay type

    Cell-based

  • Species reactivity

    Reacts with: Other species, Mammals

  • Product overview

    Cellular ROS/RNS Assay Kit (ab139473) is designed to directly monitor real time reactive oxygen and/or nitrogen species (ROS/RNS) production in live cells using fluorescence microscopy. The kit includes three fluorescent dye reagents: NO Detection Reagent (Red), Oxidative Stress Detection Reagent (Green) for total ROS detection, and Superoxide Detection Reagent (Orange). It also includes positive controls, pyocyanin and L-arginine, which are common inducers of ROS and NO production, respectively, as well as negative controls, N-acetyl-L-cysteine and c-PTIO (common scavengers of ROS and NO, respectively). Through the combination of the three detection probes with a set of specific inhibitors and activators, this kit enables discrimination among superoxide, nitric oxide and peroxynitrite.

  • Notes

    Related products

    Review the oxidative stress marker and assay guide, or the full metabolism assay guide 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.

  • Platform

    Fluorescence microscope

Properties

References (2)

Publishing research using ab139473? Please let us know so that we can cite the reference in this datasheet.

ab139473 has been referenced in 2 publications.

  • Liu B  et al. JS-K, a nitric oxide donor, induces autophagy as a complementary mechanism inhibiting ovarian cancer. BMC Cancer 19:645 (2019). PubMed: 31262254
  • Li H  et al. Evaluation of the protective potential of brain microvascular endothelial cell autophagy on blood-brain barrier integrity during experimental cerebral ischemia-reperfusion injury. Transl Stroke Res 5:618-26 (2014). PubMed: 25070048

Customer reviews and Q&As

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Cellular ROS/RNS

 Average Average 3/5 (Ease of Use)
Abreviews
Our study was focused on the mechanism of bacterial death due to plasma corona discharge exposure.
We wanted to examine the appearance of ROS and RNS.
The cells: bacterial suspension (Agrobacterium)
The concentration 1 x 105 cells/ml
The protocol was not clear. We did not understand when to add the detection MIX.
We have done as follow: 1- centrifuged the cell: 10,000g for 10 min because we are doing the experiment on bacteria 2- The positive and negative controls (untreated cells) were incubated for 1.5 hours in the detection MIX followed by addition of the inducers and inhibitors and then the cells were incubated for another 0.5 hour.
The disappointing result was that there was no difference between the ROS inducer and inhibitor as well as between the NO inducer and inhibitor.

Mrs. Yulia Lazra

Verified customer

Submitted Jul 01 2020

Question



I need some kit for my experimental work like Lipid peroxidation, NADPH oxidase & Reactive oxygen spices. in which these all will work on rat or not.because i will do my work on rat and will take their blood and tissue homogenate for further study of these parameters. these all kits having maximum 1 year expiry or not.

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Answer



I double checked and unfortunately, have confirm that we do not have a kit for measuring NADPH oxidase in our catalog at the moment. We do offer ab65349 which is a NADP/NADPH Assay Kit:

https://www.abcam.com/index.html?datasheet=65349 (or use the following: https://www.abcam.com/index.html?datasheet=65349).

I can recommend the following products

ab118970 (Lipid Peroxidation (MDA) Assay Kit)

https://www.abcam.com/index.html?datasheet=118970 (or use the following: https://www.abcam.com/index.html?datasheet=118970).

ab139473 (Cellular ROS/RNS Detection Assay Kit)

https://www.abcam.com/index.html?datasheet=139473 (or use the following: https://www.abcam.com/index.html?datasheet=139473).

ab139476 (Cellular ROS/Superoxide Detection Assay Kit)

https://www.abcam.com/index.html?datasheet=139476 (or use the following: https://www.abcam.com/index.html?datasheet=139476).

ab139473, Cellular ROS/RNS Detection Assay Kit for microscopy is used for monitoring reactive oxygen and/or nitrogen species (ROS/RNS) in live cells.

This kit can be used for discriminates among superoxide, nitric oxide and peroxynitrite by fluorescence microscopy. The kit includes three fluorescent dye reagents.

ab139476, Cellular ROS/Superoxide Detection Assay Kit: Total ROS/Superoxide detection kit is designed to directly monitor real time reactive oxygen species (ROS) production in live cells using fluorescence microscopy or flow cytometry. The kit includes two fluorescent dyes as major components: Oxidative Stress Detection Reagent (Green) for total ROS detection reagent and Superoxide Detection Reagent (Orange).

This kit distinguishes between different reactive species, such as hydrogen peroxide, peroxynitrite and hydroxyl radicals.

I can confirm that all our kits are covered by our Abpromise guarantee for m12 month if they are unopened and stored as instructed in the protocol booklet. Please note that some reagents in the kits can only be stored short term if they are opened or reconstituted.

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Question

My sample is moving cells - spermatozoa. So it is not easy to take a good picture without fixing the sample. Are the three dyes compatible with fixation of the cells (i.e, 4% PFA or other)? Would the microscopic observation work with fixed cells to reflect the changes of ROS (or RNS) levels?

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Answer

The lab says fixation will cause the dyes to leak from the cells. I think the assumption is that fixation may damage cell membranes. I did find a paper which describes fixation using methanol or acetic acid, after staining with a DCFDA derivative. DCFDA is the reagent that stains ROS in the kit ab113851, and I suspect that it is the green dye in the other two kits, the ROS/NOS and superoxide assays, though our source for those kits has not confirmed. A modified fixed staining method for the simultaneous measurement of reactive oxygen species and oxidative responses. Shen et al, 2013. PMID: 23178299 http://www.ncbi.nlm.nih.gov/pubmed/23178299

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