Prostaglandin e2 express eia kit

EM1 cells were treated with alamethicin, ionomycin, nifedipine, verapamil, or dantrolene for 1 h and then with forskolin or db-cAMP for 48 h. The culture medium (500 μl) was centrifuged at 10,000 x g at 4°C for 10 min, and the concentration of PGE2 in the supernatant was determined using a ELISA kit (Prostaglandin E2 Express EIA kit, Cayman Chemical Company, Ann Arbor, MI). One hundred μl of the supernatant was diluted 2-fold with EIA buffer for each sample measurement. Three independent sets of experiments were performed in triplicate. For IGFBP measurement at protein levels, the culture medium was centrifuged as above and IGFBP-1 in the supernatant was determined using a commercially available sandwich ELISA kit (human IGFBP-1 DuoSet kit, R&B Systems, Inc., Minneapolis, MN), as described before [11 (link)]. The concentration of IGFBP-1 was normalized to the amount of total cell protein.

The concentration of PGE2 in the culture medium was determined using a Prostaglandin E2 Express EIA kit (Cayman Chemical Co., Ann Arbor, MI) in accordance with the manufacturer’s instructions. Levels of PGE2 secreted by RAW264.7 or 3T3-L1 cells cultured separately were determined as a control.