E el m3012

Plasma insulin (E-EL-M1382c, Elabscience, Wuhan, China), GLP-1 (E-EL-M3012, Elabscience), TNF-α (RK00027, ABclonal, Wuhan, China), IL-6 (RK00008, ABclonal), and IL-4 (RK00036, ABclonal) levels were determined via ELISA according to the manufacture’s protocols. Total cholesterol (A111-1-1), triglyceride (A110-1-1), low-density lipoprotein cholesterol (A113-1-1), and high-density lipoprotein cholesterol (A112-1-1) were measured using relative commercial colorimetric enzymatic assay kits from Najing Jiancheng Bioengineering, as described previously [25 (link)].

ELISA kits for mouse ghrelin (E-EL-M0551c; Elabscience), insulin (E-EL-M1382c, Elabscience), GLP-1 (E-EL-M3012; Elabscience), and PYY (E-EL-M2375c; Elabscience) were purchased from BGsciences Biotechnology Co., Ltd. (Guangzhou, China). ELISA was performed using serum samples. Blood samples were obtained from heart and allowed to clot for 30 min at 37 °C. After centrifugation at 2000g for 15 min, the serum was collected from the blood samples. Serum was used for ELISA. The ELISA was performed according to the manufacturer’s instructions. The standard curves were plotted for each experiment.

GLP-1 and DPP IV expression in the mouse and human serum and colon was evaluated by the enzymatic immunoassay kit (competitive ELISA cat no. E-EL-H6025 (sensitivity 0.94 pg/mL) for GLP-1 and DPP IV cat no. E-EL-H0058 (sensitivity 0.19 ng/mL) for humans and GLP-1 cat no. E-EL-M3012 (sensitivity 0.10 ng/mL) and DPP IV cat no. E-EL-M2440 (sensitivity 37.50 pg/mL), Elabscience, Wuhan, China. No significant cross-reactivity in interference between mouse/human GLP-1 or DPP IV and analogues. Shortly, mouse colon samples were rinsed in ice-cold PBS to remove excess blood and feaces and weighed before the homogenization. Next, tissues were minced using motor cordless tissue grinder (Fisher scientific, Goteborg, Sweden) in the 20 volumes of ice-cold PBS. Subsequently, homogenates and serum were centrifuged for 5 min at 5000 × g, 4 °C. The supernatant and serum were used for the procedure following manufacturer’s instructions. The amount of GLP-1 and DPP IV in serum and the colonic samples was determined from the standard curve prepared with the purified GLP-1 and DPP IV standard supplied with the kit. Data of GLP-1 and DPP IV in colon samples were presented as pg per mg of colon tissue.