| Assay Method Information | |
| | In Vitro Binding Autoradiography |
| Description: | Naive rats were sacrificed by decapitation, and the brain and pituitary were collected. Slide-mounted brain sections were preincubated in an assay solution containing [125I]oCRF for 2 h at room temperature. Concentration-related displacement of test compound was assessed by including the compound in the incubation solution. The adjacent sections were incubated under the same conditions in the presence of 1 uM alpha-helical CRF for defining nonspecific binding for CRF receptor. Nonspecific binding for CRF1 receptors was defined by 1 uM DMP696 or CP-154,526. After incubation, the sections were rinsed, dried, and then placed in cassettes against iodine-sensitive storage phosphor-imaging screens (PerkinElmer Life Sciences), and the screens were then digitally scanned with a Cyclone storage phosphor-imaging system (PerkinElmer Life Sciences). Captured storage phosphor images were analyzed with OptiQuant Acquisition and Analysis software (PerkinElmer Life Sciences). |
| Affinity data for this assay | |
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