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Invitrogen™ CellROX™ Orange Reagent, for oxidative stress detection

Product Code. 12807322
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Quantity:
5 x 50 μL
Unit Size:
50µL
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12807322 5 x 50 μL 50µL
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Product Code. 12807322 Supplier Invitrogen™ Supplier No. C10443

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Includes

Five single-use vials with enough total reagent for five 96-well plates or 100 coverslips

CellROX™ Orange Reagent is a novel fluorogenic probe for measuring oxidative stress in live cells. This cell-permeant dye is non-fluorescent while in a reduced state and exhibits bright orange fluorescence upon oxidation by reactive oxygen species (ROS), with absorption/emission maxima of ∼545/565 nm.

CellROX™ Orange Reagent is a novel fluorogenic probe for measuring oxidative stress in live cells. This cell-permeant dye is non-fluorescent while in a reduced state and exhibits bright orange fluorescence upon oxidation by reactive oxygen species (ROS), with absorption/emission maxima of ∼545/565 nm. While this reagent is not formaldehyde-fixable, it can be multiplexed with other live cell-compatible probes such as GFP, CellEvent™ Caspase 3/7 Green reagent, Image-iT™ DEAD Green, Green and Deep Red MitoTracker™ dyes, and NucBlue™ reagent.

CellROX™ Orange Reagent is compatible with various platforms, such as traditional fluorescence microscopy, high content screening (HCS), and flow cytometery. This reagent is also compatible with the Tali™ Image-based Cytometer.

In addition, CellROX™ Orange Reagent is:
• Compatible with other live cell probes, including GFP, and may be added directly to serum-containing growth media
• Provided as a stable, ready-to-use DMSO solution with a simple protocol compatible with standard workflows in fluorescence microscopy
• Provided as five single-use vials containing enough 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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Shipping Condition: Wet Ice

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Specifications

Color Orange
Format Frozen
Quantity 5 x 50 μL
Content And Storage Store at ≤–20°C. Protect from light and desiccate.

For Use With (Equipment) Imaging, HCS, Cytometer
Concentration 2.5 mM stabilized solution in DMSO
Detection Method Live Cell Imaging
Excitation/Emission 545/565 nm
Indicator Oxidative stress
Product Line CellROX
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.

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.

For Research Use Only. Not for use in diagnostic procedures.

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