| Assay Method Information | |
| | Signal Transduction Inhibitory Assay |
| Description: | The biological activity of a compound was determined by measuring the activation of Smad3/Smad4 complex which is a transcription factor showing activation induced by TGF-beta stimulation. That is, a reporter plasmid having a DNA sequence (CAGA sequence), which is activated by binding to Smad3/Smad4 complex, is linked to the upstream of the luciferase gene (luminescence enzyme) was prepared. Mink lung epithelial cells CCL64 (named as x9CAGA/CCL64 cells) stably incorporating this reporter plasmid were established. The x9CAGA/CCL64 cells were cultured in DMEM medium containing 10% FBS, penicillin (100 U/ml), streptomycin (100 g/ml), and blasticidin S (1 ug/ml). The x9CAGA/CCL64 cells were seeded in a 96 well plate at a concentration of 10000 cells/well, and cultured in a 5% CO2 incubator at 37C. The next day, the medium was changed to DMEM medium containing 0.2% FBS, and the test compound was added. |
| Affinity data for this assay | |
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