im having trouble for catalase assay by Aebi 1984 for B16 cell lysate, the result increase as time goes by instead of decreasing. i wanna ask what is the optimized ver of it and does not require any dye. Also, when harvesting the cell, we used 0.025% trypsin-EDTA to harvest the adherent cells. Is trypsin-edta bad for catalase assay and I need to use cell scrapper?
Please see the attached PDF. The method works very well in our hands. All the best.
Prof. S S katyare
Since this method measure the amount of hydrogen peroxide at 240 nm and catalase dissociate hydrogen peroxide hence OD should not increase.
Increase in OD during catalase assay is due to formation of oxygen bubles in the medium. This due to inadequate H2O2 concentration and high activity of catalase. Try to change concentrations
Thank you all for the suggestions and i will try to run again and update it later
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