Figure 1. Orphanin FQ-induced concentration-dependent stimulation of intracellular calcium mobilization in CHO-K1/OPRL1/Gα15 cells. The cells were loaded with Calcium-4 (Cat. No. R8142; Molecular Devices) prior to stimulation with OPRL1 agonist, Orphanin FQ. The intracellular calcium change was measured by FLIPRTETRA. The relative fluorescent units (RFU) were recorded and normalized to plot against the log of the cumulative doses of Orphanin FQ (mean ± SEM, n = 3). The EC₅₀ of Orphanin FQ on CHO-K1/OPRL1/Gα15 cells was 0.24 μM.

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.

Figure 2. Dose dependent stimulation of intracellular cAMP accumulation upon treatment with Orphanin FQ in CHO-K1/OPRL1/Gα15 cells. d2 acceptor fluorophore-labeled cAMP (Cat. No. 62AM4PEC; Revvity) and intracellular cAMP in CHO-K1/OPRL1/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 Orphanin FQ on CHO-K1/OPRL1/Gα15 cells was 2.82 nM.

Figure 3. Dose dependent stimulation of intracellular IP-One accumulation upon treatment with Orphanin FQ in CHO-K1/OPRL1/Gα15 cells. d2 acceptor fluorophore-labeled IP-One (Cat. No. 62IPAPEB; Revvity) and intracellular IP-One in CHO-K1/OPRL1/Gα15 cells competitively bind with Europium Cryptate-labeled anti-IP-One antibody. The FRET signal decreases as the intracellular IP-One concentration rises and was measured by plate reader (Pherastar, BMG). The EC₅₀ of Orphanin FQ on CHO-K1/OPRL1/Gα15 cells was 0.20 uM.

CHO-K1/OPRL1/Gα15 Stable Cell Line

Following the cloning of the classical opioid receptors (mu, delta and kappa), the opiate receptor like-1 (ORL1) was identified as a G-protein coupled receptor (GPCR) with 65% structure homology to the other members of the opioid family. OPRL1 is a receptor for the 17 aa neuropeptide nociceptin/orphanin FQ and may be involved in the regulation of numerous brain activities, particularly instinctive and emotional behaviors. Recently, new study results are consistent with the reported high density of ORL1 receptor mRNA in dorsal raphe nucleus and with inhibitory actions of nociceptin in cells expressing ORL1.

产品编号	M00447
规格	2 vials 大单询价
数量
价格	询价


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产品简介

Product Description	Recombinant CHO-K1 cells stably overexpress human opioid related nociceptin receptor 1 (OPRL1) 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.

培养条件

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)

图例

Figure 1. Orphanin FQ-induced concentration-dependent stimulation of intracellular calcium mobilization in CHO-K1/OPRL1/Gα15 cells. The cells were loaded with Calcium-4 (Cat. No. R8142; Molecular Devices) prior to stimulation with OPRL1 agonist, Orphanin FQ. The intracellular calcium change was measured by FLIPRTETRA. The relative fluorescent units (RFU) were recorded and normalized to plot against the log of the cumulative doses of Orphanin FQ (mean ± SEM, n = 3). The EC₅₀ of Orphanin FQ on CHO-K1/OPRL1/Gα15 cells was 0.24 μM.

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.

Figure 2. Dose dependent stimulation of intracellular cAMP accumulation upon treatment with Orphanin FQ in CHO-K1/OPRL1/Gα15 cells. d2 acceptor fluorophore-labeled cAMP (Cat. No. 62AM4PEC; Revvity) and intracellular cAMP in CHO-K1/OPRL1/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 Orphanin FQ on CHO-K1/OPRL1/Gα15 cells was 2.82 nM.

Figure 3. Dose dependent stimulation of intracellular IP-One accumulation upon treatment with Orphanin FQ in CHO-K1/OPRL1/Gα15 cells. d2 acceptor fluorophore-labeled IP-One (Cat. No. 62IPAPEB; Revvity) and intracellular IP-One in CHO-K1/OPRL1/Gα15 cells competitively bind with Europium Cryptate-labeled anti-IP-One antibody. The FRET signal decreases as the intracellular IP-One concentration rises and was measured by plate reader (Pherastar, BMG). The EC₅₀ of Orphanin FQ on CHO-K1/OPRL1/Gα15 cells was 0.20 uM.

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

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CHO-K1/OPRL1/Gα15 Stable Cell Line

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