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Samples are prepared by grinding, solvent extraction, and centrifugal purification before injection
All samples were serially diluted in assay buffer in duplicate and incubated with the cells for 30 min at room temperature (RT) before detection with the cAMP dynamic 2 HTRF assay as described previously 72
The cDNA was then fourfold diluted and used as a template in quantitative PCR with IQ SYBR Green Supermix (Bio-Rad) on a CFX96 instrument (Bio-Rad) using a 3-step protocol
As with every product, confirm the shipped batch matches its Certificate of Analysis for identity, purity and salt form
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