Peroxidase Activity Quantification Using ABTS

HRP activity was estimated following the technique reported by Sanders et al. [48 (link)], employing ABTS as a reducing substrate—similar to our previous papers [8 (link),35 (link)]. The rate of change in solution absorbance at 405 nm was monitored with an Agilent 8453 UV-visible spectrophotometer (Agilent Technologies Deutschland GmbH, Waldbronn, Germany). The HRP concentration in the quartz spectrophotometric cuvette was 10⁻⁹ M, while the concentrations of H₂O₂ and ABTS were 0.3 mM and 2.5 mM, respectively. Spectrum acquisition was started immediately after the addition of H₂O₂.

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Ivanov Y.D., Pleshakova T.O., Shumov I.D., Kozlov A.F., Valueva A.A., Ivanova I.A., Ershova M.O., Larionov D.I., Repnikov V.V., Ivanova N.D., Tatur V.Y., Stepanov I.N, & Ziborov V.S. (2021). AFM and FTIR Investigation of the Effect of Water Flow on Horseradish Peroxidase. Molecules, 26(2), 306.

Publication 2021

Agilent 8453

Abts H2o2 Quartz Spectrophotometric

Corresponding Organization : Institute of Biomedical Chemistry

Other organizations : Moscow State Academy of Veterinary Medicine and Biotechnology named after Skryabin, Joint Institute for High Temperatures

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Variable analysis

independent variables

HRP concentration in the quartz spectrophotometric cuvette

dependent variables

Rate of change in solution absorbance at 405 nm

control variables

Concentration of H2O2 (0.3 mM)
Concentration of ABTS (2.5 mM)

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