Team:Caltech/Protocols/Measuring H2O2

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Measuring H2O2


Note: Based on manufacturer's instructions.

  1. Spin down approximately 100 uL of cell culture
  2. Measure peroxide concentration using a commercial colorimetric assay following the manufacturer's instructions.
  3. After incubating the reaction at room temperature for at least 20 minutes, measure the absorbance at 595 nm on a plate reader (Spectramax M2).
  4. Calculate the concentration of hydrogen peroxide by reference to a standard curve (100uM, 33.3uM, 11.1 uM, 3.7 uM and 0.0 uM H2O2) made in the corresponding media.
    • NB: Dilute culture supernatants as appropriate to bring measurements into the assay‚Äôs linear range.
An example of a typical standard curve generated by the assay. Error bars (1SD, n=3) are too small to be seen.
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