Albumin elisa kit

Female SD rats, 10 weeks old and weighing 160–180 g, were used in the present study. C-BSA induction of membranous nephropathy (MN) was adopted with previous methods [15 (link),16 ], and the experimental scheme and grouping are shown in Figure 1D. Briefly, 50 rats received intravenous injection of C-BSA (5 mg per animal, dissolved in 0.5 mL sterile 0.01 M PBS, pH 7.4) through the tail vein every day for 14 days, and the first day of receiving c-BSA was considered as day 1. Albuminuria was determined by an albumin ELISA kit (Abcam plc, Cambridge, MA, USA). All established MN rats were randomly divided into the model group (n = 24), MMF-treated group (n = 8), and losartan-treated group (n = 8). MMF (20 mg kg⁻¹) and losartan (20 mg kg⁻¹) were orally given daily to evaluate whether sNGAL could predict the treatment effect and prognosis of c-BSA-induced MN. Another 24 animals were used as sham control and received PBS i.v. injection following the same protocol. Animals from sham and MN model groups were randomly divided into three subgroups to collect blood and kidney tissues (n = 8 for each subgroup). At day 10, day 30, and day 50 after c-BSA i.v. injection, blood and kidney tissues from sham and MN subgroups were obtained. The animals in MMF-treated and losartan-treated groups were euthanized at day 50 to obtain blood and kidney tissues.

The levels of β2M and FLC were determined using the commercially available equipment Immulite 2000 XPi (Siemens Healthcare Diagnostics SA, Zürich, Switzerland; reference interval, 0.61–2.37 μg/mL) and SPA Plus® (Binding Site, Birmingham, UK; reference interval, 5.71–26.30 mg/L). The level of FGF-23 was estimated via enzymatic measurement using the FGF-23 (C-terminal) ELISA kit (Biomedica, Vienna, Austria). The albumin levels in the spent dialysates were assessed using the albumin ELISA kit (ab227933; Abcam, Cambridge, MA, USA). All serum, plasma, and spent dialysate samples were collected and sent to laboratory facilities under standardised conditions.