Myeloperoxidase Activity (MPO) Assay

The MPO activity was measured to evaluate the infiltration of neutrophils in the renal tissue due to cisplatin administration. The MPO activity in the tissue homogenate was measured according to the protocol as described elsewhere (Sahu et al., 2014). Briefly, 100 mg of the tissue was homogenized in 50 mM phosphate buffer (pH 6, 4°C) containing 0.5% hexadecyltrimethylammonium bromide. The resulting homogenate was then subjected to freeze-thaw cycles for three times and was finally centrifuged for 10 min at 30000 rpm, 4°C. MPO activity was measured by quantifying the peroxide dependent oxidation of o-dianisidine. The reaction mixture was then incubated at 37°C for 10 min. Finally, the spectrophotometric absorbance was noted at 460 nm.