E el h0058

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.