
Figure 1. CRH-induced concentration-dependent stimulation of intracellular calcium mobilization in CHO-K1/CRF1/Gα15 cells. The cells were loaded with Calcium-4 prior to being stimulated with agonist CRH. The intracellular calcium change was normalized and measured by FLIPR. The relative fluorescent units (RFU) were normalized and plotted against the log of the cumulative doses of CRH (Mean ± SEM, n = 3). The EC50 of CRH on this cell was 13.19 nM.
Notes:
EC50 value is calculated with four parameter logistic equation:
Y=Bottom + (Top-Bottom) / (1+10^((LogEC50-X)*Hill Slope))
X is the logarithm of concentration. Y is the response
Y is % stimulation of RFU and starts at Bottom and goes to Top with a sigmoid shape.

Figure 2. Dose dependent stimulation of intracellular cAMP accumulation upon treatment with urocortin in CHO-K1/CRF1/Gα15 cells. d2 acceptor fluorophore -labeled cAMP (Cat. No. 62AM4PEC; Revvity) and intracellular cAMP in CHO-K1/CRF1/Gα15 cells competitively bind with Europium Cryptate-labeled anti-cAMP monoclonal antibody. The FRET signal decreases as the intracellular cAMP concentration rises and was measured by plate reader (Pherastar, BMG). The EC50 of urocortin on CHO-K1/CRF1/Gα15 cells was 0.14 nM.
CHO-K1/CRF1/Gα15 Stable Cell Line
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