Figure 2. Dose dependent stimulation of intracellular cAMP accumulation upon treatment with melatonin in CHO-K1/MT2/Gα15 cells. d2 acceptor fluorophore -labeled cAMP (Cat. No. 62AM4PEC; Revvity) and intracellular cAMP in CHO-K1/MT2/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 melatonin on CHO-K1/MT2/Gα15 cells was 0.89 nM.

Figure 1. Melatonin-induced concentration-dependent stimulation of intracellular calcium mobilization in CHO-K1/MT2/Gα15 cells. The cells were loaded with Calcium-4 (Cat. No. R8142; Molecular Devices) prior to stimulation with MT2 agonist, melatonin. The intracellular calcium change was measured by FLIPR^TETRA. The relative fluorescent units (RFU) were recorded and normalized to plot against the log of the cumulative doses of melatonin (mean ± SEM, n = 3). The EC₅₀ of melatonin on CHO-K1/MT2/Gα15 cells was 12.75 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 along a sigmoid curve.

CHO-K1/MT2/Gα15 Stable Cell Line

Recombinant CHO-K1 cells stably overexpress human melatonin receptor 1B (MT2) on the surface and contain high levels of G protein Gαi to couple with the receptor in downstream signaling pathways.

產品編號	M00312
規格	2 vials 大單詢價
數量
價格	詢價


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產品簡介

Product Description	Recombinant CHO-K1 cells stably overexpress human melatonin receptor 1B (MT2) on the surface and contain high levels of G protein Gαi to couple with the receptor in downstream signaling pathways.
Culture Properties	Adherent
Stability	Stable through more than 16 passages with no significant changes in assay performance or expression profile.
Size	Two vials of frozen cells (>1×10⁶ per vial in 1 mL)
Storage	Store cells in liquid nitrogen immediately upon receipt. Thaw and recover cells within one year from the date received.

培養條件

Culture Medium	Ham’s F-12K (Kaighn’s), 10% FBS, 400 μg/ml Geneticin (Cat. No. 10131-035, Life Technologies), 100 μg/ml Hygromycin B (Cat. No. 10687010, Life Technologies)
Complete Growth Medium	Ham’s F-12K (Kaighn’s), 10% FBS
Freeze Medium-DATA	45% Ham’s F-12K (Kaighn’s) (Cat. No. 21127, Life Technologies), 45% FBS (Cat. No. 10099-141, Life Technologies), 10% DMSO (Cat. No. D2650, Sigma)

圖例

Figure 2. Dose dependent stimulation of intracellular cAMP accumulation upon treatment with melatonin in CHO-K1/MT2/Gα15 cells. d2 acceptor fluorophore -labeled cAMP (Cat. No. 62AM4PEC; Revvity) and intracellular cAMP in CHO-K1/MT2/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 melatonin on CHO-K1/MT2/Gα15 cells was 0.89 nM.

Figure 1. Melatonin-induced concentration-dependent stimulation of intracellular calcium mobilization in CHO-K1/MT2/Gα15 cells. The cells were loaded with Calcium-4 (Cat. No. R8142; Molecular Devices) prior to stimulation with MT2 agonist, melatonin. The intracellular calcium change was measured by FLIPR^TETRA. The relative fluorescent units (RFU) were recorded and normalized to plot against the log of the cumulative doses of melatonin (mean ± SEM, n = 3). The EC₅₀ of melatonin on CHO-K1/MT2/Gα15 cells was 12.75 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 along a sigmoid curve.

For research use only. Not intended for human or animal clinical trials, therapeutic or diagnostic use.

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CHO-K1/MT2/Gα15 Stable Cell Line

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