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Invitrogen™ OxyBURST™ Green H2DCFDA, SE

Catalog No. D2935
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D2935 5 mg
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Catalog No. D2935 Supplier Invitrogen™ Supplier No. D2935
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Description

Amine-reactive assay reagent

OxyBURST™ Green H2DCFDA succinimidyl ester (2',7'-dichlorodihydrofluorescein diacetate, SE) can be used to prepare oxidation-sensitive conjugates of wide variety of biomolecules and particles, including antibodies, antigens, peptides, proteins, dextrans, bacteria, yeast and polystyrene microspheres. Following conjugation to amines, two acetates of OxyBURST™ Green H2DCFDA can be removed by treatment with hydroxylamine at neutral pH to yield dihydrofluorescein conjugate. OxyBURST™ Green H2DCFDA conjugates are nonfluorescent until they are oxidized to corresponding fluorescein derivatives.

  • Ex/Em: approx. 492 - 495/517–527nm
  • Product is air sensitive (store under dry argon or nitrogen)
  • Product may be dissolved in DMSO, DMF, or ethanol for use
  • Indicator is cell permeant
  • Fluorescence can be monitored using a flow cytometer, fluorometer, microplate reader, or fluorescence microscope, using excitation sources and filters appropriate for fluorescein

Refer to the Molecular Probes™ Handbook for additional product information.

Cell Analysis, Cell Metabolism, Cell Viability, Proliferation and Function, Endocytosis, Exocytosis and Phagocytosis, Nitro-Oxidative Stress, Reactive Oxygen Species

Order Info

Shipping Condition: Room Temperature

Specifications

Quantity 5 mg
Product Type ROS Indicator

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).
I labeled my cell with CM-H2DCFDA for reactive oxygen detection, but upon illuminating the cell there is a significant increase in fluorescence in the control cells. Why?

If the cell is overloaded with dye, the high intracellular concentration of the dye may lead to dye-dye quenching. Upon illumination, photobleaching will occur, which will reduce the dye-dye quenching and actually increase the fluorescence (for a while, but then it will start decreasing). To solve the problem, reduce the concentration and incubation time, and try a range of incubation times and concentrations.
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.

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