Anti-Myeloperoxidase antibody [2C7] (ab25989)

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The paraffin-embedded kidney tissue sections were dewaxed by conventional paraffin and citrate buffer solution was used for antigen retrieval, followed by incubation at room temperature for 10 min using 0.3% hydrogen peroxide and the removal of the endogenous peroxidase. Next, the sections were closed by serum and placed in a humidity chamber at 37°C for 30 min. Subsequently the serum was eliminated and MPO primary antibody (diluted 1: 1000; ab25989; Abcam Company, Burlingame, CA, USA) was added. The humidity chamber with plastic bag was placed in a 4°C refrigerator overnight (> 16 h), after which the secondary antibody was added and the incubation at room temperature for 25 min was performed. The sections were then colored in diaminobenzidine (DAB), dehydrated gradually in a conventional way, transparentized, sealed with neutral resin and observed under the microscope. The number of positive MPO neutrophils in each group was calculated by randomly selecting ten fields of view at high magnification.

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