Enzyme linked immunosorbent assay elisa kit

Female Wistar rats (n = 40, body weight 220 ± 10 g, mean age 12 weeks) were obtained from the animal experimental center of Guangxi Medical University (China). Animals were housed at 20°C–25°C and 50 ± 5% humidity with ad libitum access to food and water and a 12:12 hr light/dark cycle. This study was approved by the Medical Ethics Committee of First Affiliated Hospital of Guangxi Medical University. The animals were prepared for isolation of spleen lymphocytes and preparation of prednisone‐containing serum.
Prednisone was purchased from Xianju pharmaceutical factory (Zhejiang, China). The anti‐CD3 antibody, IL‐6, IL‐23, and TGF‐β were purchased from eBioscience (California, USA). Culture medium and fetal bovine serum were purchased from Gibco (California, USA). PrimeScript RT reagent kit was purchased from Baosheng (Dalian, China). All PCR primers were designed to flank the exon–intron boundaries to avoid amplification of genomic sequences and synthesized by Huada (Beijing, China). The antibody for flow cytometry, PerCP‐Cy5.5‐conjugated anti‐CD4, phycoerythrin (PE)‐conjugated anti‐human IL‐17, APC‐conjugated anti‐CD25, and Alexa Fluor488‐conjugated anti‐CD127 were provided by Becton Dickinson (New Jersey, USA). Enzyme‐linked immunosorbent assay (ELISA) kits were purchased from Cusabio (Wuhan, China).

KIM-1 and NGAL were assessed using Enzyme-Linked Immunosorbent Assay (ELISA) kits (Cusabio Technology LLC, Wuhan, China). As specified by the manufacturer, inter-assay precision and intra-assay precision values for the KIM-1 and NGAL ELISA kits were CV% < 10% and CV% < 8%, respectively^{31 (link)}. Urine samples were analyzed for creatinine and microalbumin using an automated biochemistry analyzer (HumaStar 100; Human mbH, Wiesbaden, Germany).