| Assay Method Information | |
| | ELISA-based Correlate-EIA Direct cAMP Assay |
| Description: | INS-1E insulinoma cells were incubated in 2.5 mM glucose Krebs-Ringer buffer (pH 7.5) supplemented with 2 mM sodium bicarbonate, 10 mM HEPES, and 0.1% BSA for 2 h before start of the experiment. At time zero for each experiment, media was switched to Krebs-Ringer buffer containing 2.5 mM glucose or 16 mM glucose in the presence of 500 μM IBMX and inhibitor at the shown concentrations. After 10 min cells were lysed in 200 μl 0.1 M HCl. Intracellular cAMP contents were determined using Correlate-EIA Direct Assay (Assay Designs, Inc). |
| Affinity data for this assay | |
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