Total ROS/Superoxide detection kit for microscopy, flow cytometry and microplates

Directly Monitor Global Levels of Reactive Oxygen Species (ROS), and Specifically Superoxide



ENZ-51010	1 Kit	201.00 USD

Distinguishes between different reactive species, such as hydrogen peroxide, peroxynitrite and hydroxyl radicals
High sensitivity, specificity and accuracy for live cell studies
Compatible with major components of tissue culture media (phenol red, FBS and BSA)
Complete set of reagents, including ROS inducers and scavengers

Enzo Life Sciences’ 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). 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 reactive oxygen species (ROS), and specifically superoxide in living cells.

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Figure 1: Profiling of reactive oxygen species formation by fluorescence microscopy was achieved in HeLa cells loaded with ROS/Superoxide detection reagents 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 Specification

Quantity:	200 fluorescence microscopy assays or 50 flow cytometry assays.

Quality Control:	A sample from each lot of Total ROS/Superoxide Detection Kit is used to stain HeLa cells using the procedures described in the user manual. The stained cells are analyzed using a wide-field fluorescence microscope equipped with standard green (490/525 nm) and orange (550/620 nm) filter set. The following results are obtained: - ROS positive control samples induced with Pyocyanin exhibit bright orange signal in nucleus as well as bright green fluorescence in the cytoplasm. - Cells pretreated with the ROS inhibitor don’t demonstrate any green or orange fluorescence signal upon induction.

Kit/Set Contains:	Oxidative Stress Detection Reagent (Green), 300 nmoles Superoxide Detection Reagent (Orange), 300 nmoles ROS Inducer (Pyocyanin), 1 µmole ROS Inhibitor (N-acetyl-L-cysteine), 2 x 10 mg Wash Buffer Salts, 1 pack

Application:	This kit is designed to directly monitor real time reactive oxygen and/or nitrogen species (ROS/RNS) production in live cells using fluorescence microscopy and/or flow cytometry.

Short Term Storage:	-20°C

Long Term Storage:	-80°C

Use/Stability:	With proper storage, the kit components are stable up to the date noted on the product label. Store kit at -20°C in a non-frost free freezer, or -80°C for longer term storage.

Background / Technical Information:	The Total ROS/Superoxide Detection Kit is a member of the CELLestial^® product line, reagents and assay kits comprising fluorescent molecular probes that have been extensively benchmarked for live cell analysis applications. CELLestial^® reagents and kits are optimal for use in demanding imaging applications, such as confocal microscopy, flow cytometry and HCS, where consistency and reproducibility are required.

General Literature References

Fluorescent and luminescent probes for measurement of oxidative and nitrosative species in cells and tissues: progress, pitfalls, and prospects: P. Wardman; Free Radic. Biol. Med. 43, 995 (2007), Abstract;

Fluorescence probes used for detection of reactive oxygen species: A. Gomes, et al.; J. Biochem. Biophys. Methods 65, 45 (2005), Abstract;

Determination of mitochondrial reactive oxygen species: methodological aspects: C. Batandier, et al.; J. Cell. Mol. Med. 6, 175 (2002), Abstract;

Methods of detection of vascular reactive species: nitric oxide, superoxide, hydrogen peroxide, and peroxynitrite: M.M. Tarpey & I. Fridovich; Circ. Res. 89, 224 (2001), Abstract;