human APJ cyclic AMP (hcAMP) activity

16:53 Sep 29, 2018
This question was closed without grading. Reason: No acceptable answer

English to Russian translations [PRO]
Science - Biology (-tech,-chem,micro-) / биохимия�
English term or phrase: human APJ cyclic AMP (hcAMP) activity
HEK293 cells stably expressing human APJ receptor were used to assess the activity of compounds. Cultured cells were detached and resuspended in the cAMP Homogeneous Time-Resolved Fluorescence (HTRF) assay buffer. Serial dilutions of a compound together with assay buffer containing 0.2nM IBMX and 2 μM forskolin were added to each well containing 5,000 cells and incubated for 30 minutes at room temperature. Subsequently, cAMP D2 reagent was added in the lysis buffer followed by the EuK antibody and incubated for 60 min. The fluorescence emission ratio was measured using fluorometer. The intracellular cAMP concentrations (compound-stimulated inhibition of forskolin-mediated cAMP production) were calculated by extrapolation from a standard curve using known cAMP concentrations. The EC50 values were obtained by fitting the data to a sigmoidal concentration-response curve with variable slope. The maximal achievable inhibition of forskolin-induced cAMP levels (Ymax) for each compound was expressed as relative percentage of inhibition attained using pyroglutamated apelin-13 ((Pyr1)apelin-13) peptide, which was set to 100%.
The examples disclosed below were tested in the APJ in vitro assays described above and were found having ***human APJ cyclic AMP (hcAMP) activity***.
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