By Lester Packer, Enrique Cadenas

This new quantity of Methods in Enzymology maintains the legacy of this most effective serial with caliber chapters authored by way of leaders within the box. this is often the second one of 3 volumes on hydrogen peroxide and mobile signaling, and comprises chapters on such themes because the mobile steady-state of H2O2, evaluating peroxiredoxin sensitivity in the direction of inactivation by way of peroxide substrates, and peroxiredoxins as preferential goals in H2O2-induced signaling.

  • Continues the legacy of this most desirable serial with caliber chapters authored through leaders within the box
  • Covers hydrogen peroxide and cellphone signaling
  • Contains chapters on such issues as the mobile steady-state of H2O2, evaluating peroxiredoxin sensitivity in the direction of inactivation through peroxide substrates, and peroxiredoxins as preferential goals in H2O2-induced signaling

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Extra info for Hydrogen Peroxide and cell signaling, Part B

Sample text

KGPxPostNuclear cannot be compared directly with kintact cells or kcatalase because the yield of the extraction of the postnuclear cell fraction is not 100%. So, an additional measurement is needed, namely, the determination of catalase activity in the postnuclear protein extract used for determination of GPx activity (kcatalasePostNuclear), and instead of Eq. 4), Eq. 11) is used. R¼ kcatabolism kGPxPostNuclear ðkcatalase =kcatalasePostNuclear Þ þ kcatalase ¼ kintact cells kintact cells ð1:11Þ The protocol described here was optimized for MCF-7 cells.

GSH 33 mM in phosphate buffer (can be stored at À20  C). 5. 5% NaHCO3). 6. 35 mM (in water prepared daily). 7. Triton X-100 10% (v/v). , 1979). 8 Â 106 MCF-7 cells). Preincubate at 37  C for 10 min and then add 35 mM H2O2 to the cuvette to start the reaction. NADPH consumption is followed at 340 nm (e ¼ 6220 MÀ1 cmÀ1). 2 s) until well after all peroxide is consumed (the rate of reaction decreases rapidly when all peroxide is consumed). It is important to continue to record the absorbance after this because the rate of H2O2 Gradients 15 NADPH oxidation in the absence of peroxide is needed to analyse the data.

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