Mouse cxcl1 kc duoset elisa

Mesothelial cells from C57BL/6 WT and NOD1-KO mice were prepared from the peritoneum and external surface of the liver, spleen, and kidneys of adult mice as described (Park et al., 2007 (link)). Briefly, pieces of the peritoneum and intact organs were obtained from sacrificed mice and digested with 0.25%-trypsin-EDTA solution for 50 min at 37°C. Intact tissues and tissue debris were discarded and the cell suspension was centrifuged at 1000 rpm for 5 min. The pellet was resuspended in Dulbecco's modified eagle medium (DMEM) supplemented with 15% heat-inactivated FBS and 1% P/S and cultured overnight. The next day, non-adherent cells were removed and the resultant mesothelial cells were grown in the above culture medium for 1 week. Mesothelial cells were used between passages 2 and 4 for the stimulatory assay. The cells were stimulated with heat-inactivated bacteria at a bacteria/mesothelail cells ratio of ~10:1, 1:1 and 0.1 for 12h. Culture supernatants were collected and assayed for CXCL1 production by mouse CXCL1/KC DuoSet ELISA (R&D Systems, DY453).