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. Author manuscript; available in PMC: 2015 Feb 1.
Published in final edited form as: Biochim Biophys Acta. 2013 May 7;1840(2):10.1016/j.bbagen.2013.04.040. doi: 10.1016/j.bbagen.2013.04.040

TABLE.

Attributes of an ideal probe for O2· and H2O2

  1. Responsive to low concentrations of the oxidant of interest

  2. Sensitive to relevant concentrations of oxidant

  3. Responds to oxidant more rapidly than do competing reactants

  4. Specific and selective for the oxidant of interest

  5. Quantitative

  6. Permeable to cells & can be modified to target specific organelles

  7. Nontoxic to cells and does not perturb cellular redox state

  8. Chemistry well-defined and stoichiometry of reaction with oxidant of interest known

  9. Product nonreactive; no secondary reactions & no secondary signals

  10. Easy to use, requiring neither special expertise nor dedicated equipment

Some properties (e.g. 6 and 7) would be relevant only when monitoring O2· and H2O2 in intracellular compartments. The relative importance of specific attributes depends on the goal of the study, whether it is to quantitate precisely the products of NADPH oxidase activity, to identify cellular compartments in which oxidants are generated, or to monitor kinetics of oxidant production in the context of signaling cascades.