Hello,
I want to perform a quantitative Peroxide assay with plant extracts using the Pierce Kit (Pierce™ Quantitative Peroxide Assay Kit (Aqueous), #23280). I already did some preliminary test with PBS based cell extracts but the obtained values are usually very low. Hence my question would be if there is a good way to inhibit Catalases and/or other Peroxide metabolizing enzymes to prevent decomposition of H2O2 during the procedure?
Thanks in advance!
Sodium azide might do the trick. (If the peroxide assay is not based on peroxidases).
Alternatively, you could try hevay metals, e.g. copper sulphate (but not in conjunction with azide, as copper and other heavy metals will form highly explosive azides).
Hi, thanks a lot for the hint!
Any recommendation for a working concentration, or a publication to check? The assay works with Xylene Orange and the oxidation of Fe2+ ions.
I will check if that works.
best
Sebastian
If you mix Fe2+ and Cu2+, there might be oxidation to Fe3+ and precipitation of Cu(I). Check the redox potentials (or just give it a try, and use defined compositions w/o lysates as controls and for calibration).
Not sure if hydrogen peroxide survives an increase in temperature, but if so, denaturing catalase(s) at elevated temperature also might help.
I agree with Wolfgang Schechinger. Azide should do the job. I used it to quench the activity of endogenous peroxidases in Western Blot application. The working concentration for this purpose is 1 mM according to this publication: Article Reutilization of Immunoblots after Chemiluminescent Detection
All the best,
Andreas
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