Immunohistochemical Analysis of Kidney Sections

Kidney sections (4 µm) were deparaffinized and rehydrated. Endogenous peroxidase activity was blocked using 0.3% H₂O₂ at 37 °C for 10 min. Heat-mediated antigen retrieval was performed using 10% citrate buffer. Then the sections were incubated with primary antibodies followed by secondary antibody and visualized with diaminobenzydine (DAB). Primary antibodies and dilutions used were as follows: Col IV (1:200; ab6586, Abcam), SNRK (1:500; ab96762, Abcam). The stained slides were photographed using an OLYMPUS BX51 microscope and the percentages of positive cells and staining intensities were scored as previously described^{58 (link)}.