Pai 1 total mouse elisa kit

At 5 days after STZ injection and during week 15 of the HFD, mice were fasted overnight (12 h) and blood samples were collected by cutting the tip of the tail. Fasting serum glucose levels were measured using an Accu-Chek blood glucose meter (Roche Diagnostics, Castle Hill, Australia). The levels of serum cholesterol, triglycerides, and blood urea nitrogen (BUN) were determined in serum with an Idexx VetTest Biochemical Analyzer (Westbrook, ME, USA). The serum PAI-1 levels, which include active, inactive, and latent forms of PAI-1, were assessed using a PAI-1 Total Mouse ELISA kit (Abcam) according to the manufacturer’s instructions.

After collecting blood, we separated plasma and serum by centrifugation and stored them at −80 °C prior to analysis. The uPA was measured by using the Mouse uPA Total Antigen Assay enzyme-linked immunosorbent assay (ELISA) kit (Molecular Innovations, Novi, MI, USA), with the intra- and inter-assay coefficients of variation being 6.18% and 7.47%, respectively. The soluble uPAR were measured by using the Mouse uPAR DuoSet ELISA kit (DY531, R&D system, Minneapolis, MN, USA). The mean coefficient of variation in these assays was 5%. Murine PAI-1 was measured by using the PAI-1 Total Mouse ELISA kit (ab157529, Abcam, Cambridge, MA, USA) with the intra- and inter-assay coefficients of variation being 7.9% and 12.9%, respectively. Total cholesterol, TG, serum and urine sodium, and creatinine were analyzed by spectrophotometry (Fuji Dri-Chem 3000, Fuji Film, Kanagawa, Japan). The FENa was applied to evaluate renal filtration and calculated by the ratio of urine and plasma sodium and creatinine. Serum insulin was measured using the Mouse Insulin ELISA Kit (Mercodia AB, Uppsala, Sweden) with the intra- and inter-assay coefficients of variation being 3.4% and 3.6%, respectively. All samples were assayed in duplicate. HOMA-IR was calculated to assess insulin resistance [36 (link)].