CHO-K1/ADORA1/Gα15 Stable Cell Line

Figure 1. NECA-induced concentration-dependent stimulation of intracellular calcium mobilization in CHO-K1/ADORA1/Gα15 cells. The cells were loaded with Calcium-4 (Cat. No. R8142; Molecular Devices) prior to stimulation with ADORA1 agonist, NECA (Cat. No. HY-103173, MCE). The intracellular calcium change was measured by FLIPRTETRA. The relative fluorescent units (RFU) were recorded and plotted against the log of the cumulative doses of NECA (mean ± SEM, n = 3). The EC₅₀ of NECA on CHO-K1/ADORA1/Gα15 cells was 0.12 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 along a sigmoid curve.

Figure 2. Dose dependent increase of intracellular cAMP accumulation upon treatment with CGS-15943 in CHO-K1/ADORA1/Gα15 cells. d2 acceptor fluorophore-labeled cAMP (Cat. No. 62AM4PEC; Revvity) and intracellular cAMP in CHO-K1/ADORA1/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 (Varioskan, Thermo Fisher). The EC₅₀ of CGS-15943 on CHO-K1/ADORA1/Gα15 cells was 67.83 nM.

Recombinant CHO-K1 cells stably overexpress human adenosine A1 receptor (ADORA1) on the surface and contain high levels of G protein Gαi to couple with the receptor in downstream signaling pathways.

产物编号	M00324
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 adenosine A1 receptor (ADORA1) 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, 200 μg/ml Zeocin (Cat. No. R250-01, Life Technologies), 200 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. NECA-induced concentration-dependent stimulation of intracellular calcium mobilization in CHO-K1/ADORA1/Gα15 cells. The cells were loaded with Calcium-4 (Cat. No. R8142; Molecular Devices) prior to stimulation with ADORA1 agonist, NECA (Cat. No. HY-103173, MCE). The intracellular calcium change was measured by FLIPRTETRA. The relative fluorescent units (RFU) were recorded and plotted against the log of the cumulative doses of NECA (mean ± SEM, n = 3). The EC₅₀ of NECA on CHO-K1/ADORA1/Gα15 cells was 0.12 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 along a sigmoid curve.

Figure 2. Dose dependent increase of intracellular cAMP accumulation upon treatment with CGS-15943 in CHO-K1/ADORA1/Gα15 cells. d2 acceptor fluorophore-labeled cAMP (Cat. No. 62AM4PEC; Revvity) and intracellular cAMP in CHO-K1/ADORA1/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 (Varioskan, Thermo Fisher). The EC₅₀ of CGS-15943 on CHO-K1/ADORA1/Gα15 cells was 67.83 nM.

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

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CHO-K1/ADORA1/Gα15 Stable Cell Line

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