CellROX™ Green Reagent, for oxidative stress detection
CellROX™ Green Reagent, for oxidative stress detection
Invitrogen™

CellROX™ Green Reagent, for oxidative stress detection

CellROX Green Reagent is a novel fluorogenic probe for measuring oxidative stress in live cells. The cell-permeant dye is weaklyRead more
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Catalog number C10444
Price (USD)
626.00
Each
Add to cart
Price (USD)
626.00
Each
Add to cart
CellROX Green Reagent is a novel fluorogenic probe for measuring oxidative stress in live cells. The cell-permeant dye is weakly fluorescent while in a reduced state and exhibits bright green photostable fluorescence upon oxidation by reactive oxygen species (ROS) and subsequent binding to DNA, with absorption/emission maxima of ∼ 485/520 nm. This reagent can be formaldehyde-fixed and its signal survives detergent treatment, which makes it multiplexable with other compatible dyes and antibodies.

CellROX Green Reagent is compatible with various platforms, such as traditional fluorescence microscopy, high content screening (HCS), flow cytometery, and microplate-based fluorimetry or high throughput screening (HTS). This reagent is also compatible with various benchtop instruments such as our FLoid, Tali, and Attune instruments.

In addition, CellROX Green Reagent is:

• Compatible with other live cell dyes, antibodies, and in cells expressing RFP
• Provided as a stable, ready-to-use DMSO solution with a simple no-wash protocol compatible with standard workflows in fluorescence microscopy
• Provided as five single-use vials containing sufficient total reagent for five 96-well plates or 100 coverslips

Oxidative stress results from an imbalance between the production of reactive oxygen species (ROS) and the ability of cells to scavenge them. ROS play an important role in the progression of several diseases, including inflammation, atherosclerosis, aging, and age-related degenerative disorders.

For Research Use Only. Not intended for any animal or human therapeutic or diagnostic use.

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For Research Use Only. Not for use in diagnostic procedures.
Specifications
ColorGreen
FormatFrozen
Quantity5 x 50 μL
For Use With (Equipment)Imaging, HCS, Cytometer
Concentration2.5 mM stabilized solution in DMSO
Detection MethodLive Cell Imaging
Excitation/Emission485/520 nm
IndicatorOxidative stress
Product LineCellROX™
Unit SizeEach
Contents & Storage
Store at ≤–20°C. Protect from light and desiccate.

Frequently asked questions (FAQs)

I need a formaldehyde-fixable reactive oxygen species detection assay. Is H2 DCFDA fixable?

H2DCFDA and similar derivatives are not fixable. The same goes for dihydroethidium and dihydrorhodamine. However, CellROX Deep Red and CellROX Green are retained for a limited time upon fixation with formaldehyde. CellROX Green may be retained upon subsequent Triton X-100 permeabilization. Avoid the use of any acetone or alcohol-based fixatives or fixatives that include alcohol, such as formalin.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

Can CellROX Green dye diluted in DMSO be subjected to freeze-thaw cycles?

CellROX Green dye diluted in a DMSO stock solution is stable for multiple freeze-thaw cycles, but it is not stable long-term in aqueous solution. Please note, every time the stock solution is opened, some of the dye can oxidize, which will increase background noise. DMSO readily absorbs moisture from the air, especially when cold, so be sure to completely thaw the stock solution to room temperature before opening and only open briefly. Water in the DMSO will gradually cause the dye to precipitate and come out of solution. Minimize the number of times you use a stock solution to several freeze-thaw cycles or make small aliquots to reduce the number of freeze-thaw cycles.

Find additional tips, troubleshooting help, and resources within our Flow Cytometry Support Center.

What is the difference between the CellROX Green Flow Cytometry Assay Kit (Cat. No. C10492) and CellROX Green Reagent, for oxidative stress detection (Cat. No. C10444)?

The main difference between the CellROX Green Flow Cytometry Assay Kit (Cat. No. C10492) and CellROX Green Reagent, for oxidative stress detection (Cat. No. C10444), is the suggested final working concentrations and application. The CellROX Green in the CellROX Green Flow Cytometry Assay Kit is specifically designed for flow cytometry analysis and is used at a lower concentration than what is required for imaging, as fluorescence-activated cell sorting (FACS) is a very sensitive detection system.

The standalone CellROX Green Reagent (Cat. No. C10444) is marketed for fluorescent microscopy and require working concentrations appropriate for imaging. This can typically be as much as 10-fold higher than the concentration recommended for flowcytometry. The standalone CellROX Green Reagent can also be used for flowcytometry; however, the working concentration will have to be optimized. For the flow cytometry quick reference, use the link below.

CellROX Flow Cytometry Assay Kit Quick Reference

Find additional tips, troubleshooting help, and resources within our Flow Cytometry Support Center.

What dyes can I use to detect reactive oxygen species (ROS) in my bacteria?

Many dyes that are used on mammalian cells have also been shown to be useful in bacterial cells. For example, CellROX Deep Red Reagent has been shown to work in B. subtilis (see Reference: http://www-brs.ub.ruhr-uni-bochum.de/netahtml/HSS/Diss/RaatschenNadja/diss.pdf). If you are interested in a particular dye, but are not sure if it will work on your bacteria, literature searches are the best way to check to see if it has been tested. If not, then it may be worth testing yourself.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

I am trying to label my cells with a reactive oxygen species (ROS) indicator dye, but I am not seeing a significant difference in signal. What could be happening?

First, make sure you have both a negative (untreated) and positive (ROS-induced) sample to compare. A good positive control can be the use of 100 µM menadione for one hour or 50 µM nefazodone for 24 hours. H2O2 can also be used, though it does not work well for CellROX dyes. Some dyes, such as H2DCFDA, require esterase cleavage, so don't incubate in the presence of serum (which contains esterases that can prematurely cleave the dye). If your positive control does not show significant change compared to the negative control, try increasing the concentration and label time for the dye. Our manuals give starting recommendations. Be sure to image your live cells as soon as possible. Only two dyes (CellROX Green and CellROX Deep Red) are retained with formaldehyde fixation. Finally, make sure you are using filters and instrument settings to match the excitation and emission spectra of the dye.

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.