CHO-K1/OX1 Stable Cell Line

Figure 1. Orexin A-induced concentration-dependent stimulation of intracellular calcium mobilization in CHO-K1/OX1 cells. The cells were loaded with Calcium-4 (Cat. No. R8142, Molecular Devices) prior to being stimulated with agonist orexin A. The intracellular calcium change was measured by FLIPR. The relative fluorescent units (RFU) were recorded and plotted against the log of the cumulative doses of orexin A (Mean ± SEM, n = 3). The EC₅₀ of orexin A on this cell was 0.59 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 RFU and starts at Bottom and goes to Top with a sigmoid shape.

Figure 2. Dose dependent stimulation of intracellular IP-One accumulation upon treatment with orexin A in CHO-K1/OX1 cells. d2 acceptor fluorophore-labeled IP-One (Cat. No. 62IPAPEB; Revvity) and intracellular IP-One in CHO-K1/OX1 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 (Varioskan, Thermo Fisher). The EC₅₀ of orexin A on CHO-K1/OX1 cells was 12.08 nM.

Recombinant CHO-K1 cells stably overexpress human hypocretin receptor 1 (OX1) on the surface and contain high levels of G protein Gαq to couple with the receptor in downstream signaling pathways.

产物编号	M00224
规格	2 vials 大单询价
数量
价格	询价


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产物介绍

Product Description	Recombinant CHO-K1 cells stably overexpress human hypocretin receptor 1 (OX1) on the surface and contain high levels of G protein Gαq 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)
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. Orexin A-induced concentration-dependent stimulation of intracellular calcium mobilization in CHO-K1/OX1 cells. The cells were loaded with Calcium-4 (Cat. No. R8142, Molecular Devices) prior to being stimulated with agonist orexin A. The intracellular calcium change was measured by FLIPR. The relative fluorescent units (RFU) were recorded and plotted against the log of the cumulative doses of orexin A (Mean ± SEM, n = 3). The EC₅₀ of orexin A on this cell was 0.59 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 RFU and starts at Bottom and goes to Top with a sigmoid shape.

Figure 2. Dose dependent stimulation of intracellular IP-One accumulation upon treatment with orexin A in CHO-K1/OX1 cells. d2 acceptor fluorophore-labeled IP-One (Cat. No. 62IPAPEB; Revvity) and intracellular IP-One in CHO-K1/OX1 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 (Varioskan, Thermo Fisher). The EC₅₀ of orexin A on CHO-K1/OX1 cells was 12.08 nM.

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

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CHO-K1/OX1 Stable Cell Line

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