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Invitrogen™ Dihydroethidium (Hydroethidine)

Catalog No. D1168
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25 mg
10 x 1 mg
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Catalog No. Quantity
D1168 25 mg
D11347 10 x 1 mg
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Catalog No. D1168 Supplier Invitrogen™ Supplier No. D1168
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Description

Free radical sensor

  • Superoxide indicator dihydroethdium, also called hydroethidine, exhibits blue-fluorescence in cytosol until oxidized, where it intercalates within cell's DNA, staining its nucleus a bright fluorescent red
  • Excitation Emission: 518/606nm

Cat. No. D23107 packaged 5mM solution stabilized in DMSO.

Apoptosis, Cell Analysis, Cell Structure, Cell Tracing and Tracking, Cell Viability, Proliferation and Function, Free Radical Detection, Nitro-Oxidative Stress, Nucleus, Nucleoli and Nuclear Envelope, Organelle Tracing, Reactive Oxygen Species

Order Info

Shipping Condition: Room Temperature

Specifications

Quantity 25 mg
Excitation/Emission 518/605 nm

Frequently Asked Questions (FAQs)

I want to assay cells for reactive oxygen species using carboxy-H2DCFDA, but I want to do so with a plate reader instead of microscope. Will it work?

It has been done. The problem is that plate readers are less sensitive than microscopes, with far less signal-to-background difference. It is worth trying, but first optimize concentrations and loading times with control cells, use a plate with little to no autofluorescence, and possibly optimize the gain setting in order to get the best signal possible. But don't expect the same sensitivity, even with optimization.
I have GFP-transfected cells and need to label for reactive oxygen species. Can I use H2DCFDA?

This is not recommended as the two dyes overlap in the emission wavelength. There are other ROS reagents available in different wavelengths, such as CellROX Deep Red, which emits in the far-red range (665 nm), or dihydroethidium, which is emits in the visible red range (620 nm).
What is the protocol for the use of Dihydroethidium (Hydroethidine) (Cat. No. D11347)?

  • Dissolve Dihydroethidium (DHE) in high quality, anhydrous DMSO or DMF up to 10 mM and store in aliquots at -20 to -80°C under argon or nitrogen. DHE oxidizes more readily in solution and is very sensitive to light. Avoid repeated freeze/thaws.
  • Treat adherent cells to induce reactive oxygen species (ROS).
  • Wash 2-3 times with a physiological buffer (PBS, HBSS).
  • Incubate about 30 minutes at 37°C in about 1-20 µM DHE diluted in buffer.
  • Wash 2-3 times more with buffer.
  • Image or analyse using filters appropriate for excitation/emission = 302, 518/605 nm.

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