Creatinine serum detection kit

Mouse blood was harvested from one-side eyeball while the mice were in deep anesthesia and then stored at −80 °C. Serum samples were extracted by centrifuge. Biochemical parameters, aspartate aminotransferase (AST, AM102-K, Asan Pharmaceutical, Seoul, Korea), alanine aminotransferase (ALT, AM103-K), blood urea nitrogen (BUN, BUN Serum Detection Kit, ARBOR ASSAYS, Ann Arbor, MI, USA), and creatinine (Creatinine Serum Detection Kit, ARBOR ASSAYS, MI, USA) were detected according to the manufacturers’ manuals.

Plasma urea concentration was determined in plasma samples using the QuantiChrom™ Urea Assay Kit ( Bioassay Systems, CA, USA) according to manufacturer’s protocol. Plasma creatinine concentration was determined by Creatinine Serum Detection Kit (Arbor Assays, MI, USA) according to manufacturer’s protocol.

IMQ cream (5%, Mochida Pharmaceutical) was administered on the skin of the left ear of age-matched 8–9-week-old female B6 WT and Padi4 KO mice every alternate day up to 8 weeks as reported previously (13 (link)). Degree of proteinuria was evaluated semi-quantitatively using Albustix (Siemens Healthineers) every week. Serum anti-dsDNA IgG titers were measured using an anti-dsDNA antibody mouse ELISA kit (Shibayagi). Serum anti-Sm (IgG, IgA, and IgM) antibodies were detected using mouse anti-Sm Total IgG ELISA kit (Alpha Diagnostic). Matrix metalloproteinase-9 (MMP-9) concentrations in kidney supernatants were measured using a Mouse Total MMP-9 Quantikine ELISA kit (R&D Systems). Serum BUN and Creatinine was measured using QuantiChrom Urea Assay Kit (BioAssay) and Serum Creatinine Detection Kit (Arbor Assays), respectively. Renal histopathology was analyzed by hematoxylin-eosin (HE) staining and immunohistochemistry. Myeloid lineage cells in the kidneys and spleen were isolated and analyzed by flow cytometry (MoFlo XDP, Beckman Coulter).