Rat iga elisa kit

Adiponectin, IgA, and glucose were analyzed using Mouse/Rat Adiponectin ELISA Kit (Otsuka Pharmaceutical Co. Ltd., Tokyo, Japan), Rat IgA ELISA kit (Bethyl Laboratories Inc., Montgomery, TX, USA), and Glucose CII-test Wako (Wako Pure Chemical Industries Ltd., Osaka, Japan), respectively. Transaminase CII-test Wako (Wako Pure Chemical Industries) was used to determine the aspartate aminotransferase (AST)/alanine aminotransferase (ALT) activities.

Blood tests, including complete blood count, peripheral neutrophil/lymphocyte ratio (NLR), aspartate aminotransferase, alanine aminotransferase (ALT), total bilirubin (T-Bil), serum albumin, and serum creatinine, were performed in a professional clinical laboratory (Japan Clinical Laboratories, Kyoto, Japan).
To evaluate the immune function, the CD4/CD8 T-cell ratio was analyzed. The conjugated mouse anti-rat monoclonal antibodies used for flow cytometry, APC-conjugated CD3, FITC-conjugated CD4, and PE-conjugated CD8a, were commercially available (BD Biosciences, San Josè, CA, United States). Samples were acquired using a BD Accuri C6 (BD Biosciences).
The fecal IgA content was determined to evaluate intestinal barrier function by enzyme-linked immunosorbent assay (ELISA) using a rat IgA ELISA kit (Bethyl Laboratories, Inc., Montgomery, TX, United States). Serum lipopolysaccharide (LPS) levels were evaluated using a rat LPS ELISA kit (CUSABIO, Wuhan, China).

Feces were collected and stored at − 80 °C before analysis. In brief, they were dried for 30 min in a vacuum dryer at room temperature. Later, 0.1 g of a dried sample was weighed and vortexed in 1 ml of normal saline and centrifuged, and the supernatant was collected and centrifuged. The concentrations of secretory IgA in feces and sera were determined using Rat IgA ELISA Kit (E111-102; Bethyl Laboratories, Inc., Montgomery, TX, USA), in accordance with the manufacturer’s protocol.