Figure 1. Dopamine-induced concentration-dependent stimulation of intracellular calcium mobilization in CHO-K1/D2/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 9.13 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 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/D2/Gα15 cells. d2 acceptor fluorophore-labeled cAMP (Cat. No. 62AM4PEB; Revvity) and intracellular cAMP in CHO-K1/D2/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 (Pherastar, BMG). The EC₅₀ of dopamine on CHO-K1/D2/Gα15 cells was 20.50 nM.

CHO-K1/D2/Gα15 Stable Cell Line

Dopamine is the predominant catecholamine neurotransmitter found in mammalian brain, where it controls a variety of functions including locomotor activity, cognition, emotion, positive reinforcement, food intake, and endocrine regulation. It also plays multiple roles in the periphery as a modulator of cardiovascular function, catecholamine release, hormone secretion, vascular tone, renal function, and gastrointestinal motility. The dopamine receptor family consists of five members, which are classified into two groups, D1-like (D1 and D5) and D2-like (D2, D3, and D4). Dopamine receptor 2 is mainly expressed in the brain. It has splicing variants, D2L and D2S. D2R receptor is implicated in a number of neurological and psychiatric conditions. Drugs acting at dopamine D2 receptors (D2R) are commonly used to alleviate symptoms for Parkinson's disease, schizophrenia, and depression.

产品编号	M00152
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 dopamine receptor D2 (D2) 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), 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/D2/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 9.13 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 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/D2/Gα15 cells. d2 acceptor fluorophore-labeled cAMP (Cat. No. 62AM4PEB; Revvity) and intracellular cAMP in CHO-K1/D2/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 (Pherastar, BMG). The EC₅₀ of dopamine on CHO-K1/D2/Gα15 cells was 20.50 nM.

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

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CHO-K1/D2/Gα15 Stable Cell Line

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