CHO-K1/D2s/Gα15 Stable Cell Line

Figure 1. Dopamine-induced concentration-dependent stimulation of intracellular calcium mobilization in CHO-K1/D2s/Gα15 cells. The cells were loaded with Calcium-4 prior to being stimulated with agonist dopamine. 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 dopamine (Mean ± SEM, n = 3). The EC₅₀ of dopamine on this cell was 3.61 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 dopamine in CHO-K1/D2s/Gα15 cells. d2 acceptor fluorophore-labeled cAMP (Cat. No. 62AM4PEB; Revvity) and intracellular cAMP in CHO-K1/D2s/Gα15 cells competitively bind with Europium Cryptate-labeled anti-cAMP antibody. The FRET signal decreases as the intracellular cAMP concentration rises and was measured by plate reader (Varioskan, Thermo Fisher). The EC₅₀ of dopamine on CHO-K1/D2s/Gα15 cells was 5.53 nM.

Recombinant CHO-K1 cells stably overexpress human dopamine receptor D2s (D2s) on the surface and contain high levels of G protein Gαi to couple with the receptor in downstream signaling pathways. This variant encodes a 414 aa isoform (short) and is 29 aa shorter than the long one.

产物编号	M00274
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 dopamine receptor D2s (D2s) on the surface and contain high levels of G protein Gαi to couple with the receptor in downstream signaling pathways. This variant encodes a 414 aa isoform (short) and is 29 aa shorter than the long one.
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), 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. Dopamine-induced concentration-dependent stimulation of intracellular calcium mobilization in CHO-K1/D2s/Gα15 cells. The cells were loaded with Calcium-4 prior to being stimulated with agonist dopamine. 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 dopamine (Mean ± SEM, n = 3). The EC₅₀ of dopamine on this cell was 3.61 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 dopamine in CHO-K1/D2s/Gα15 cells. d2 acceptor fluorophore-labeled cAMP (Cat. No. 62AM4PEB; Revvity) and intracellular cAMP in CHO-K1/D2s/Gα15 cells competitively bind with Europium Cryptate-labeled anti-cAMP antibody. The FRET signal decreases as the intracellular cAMP concentration rises and was measured by plate reader (Varioskan, Thermo Fisher). The EC₅₀ of dopamine on CHO-K1/D2s/Gα15 cells was 5.53 nM.

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

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CHO-K1/D2s/Gα15 Stable Cell Line

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