Figure 1. Carbacol-induced concentration-dependent stimulation of intracellular calcium mobilization in CHO-K1/M1 cells. The cells were loaded with Calcium-4 prior to being stimulated with agonist carbacol. The intracellular calcium change was normalized and measured by FLIPR. The relative fluorescent units (RFU) were normalized and plotted against the log of the cumulative doses of carbacol (Mean ± SEM, n = 3). The EC₅₀ of carbacol on this cell was 30.92 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 IP-One accumulation upon treatment with carbacol in CHO-K1/M1 cells. d2 acceptor fluorophore-labeled IP-One (Cat. No. 62IPAPEB; Revvity) and intracellular IP-One in CHO-K1/M1 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 carbacol on CHO-K1/M1 cells was 1.39 μM.

CHO-K1/M1 Stable Cell Line

M1 was expressed in the CNS such as cerebral cortex, basal ganglia, limbic areas, vestibular system and esophageal smooth muscle. Synaptic transmission by muscarinic acetylcholine receptors (mAChRs) is employed throughout the central and peripheral nervous systems to elicit a large and diverse array of neurophysiological actions. An important aspect of mAChR functional diversity is reflected by the multitude of biochemical and electrophysiological actions evoked by acetylcholine binding to mAChRs, which include the regulation of intracellular levels of cAMP, cGMP and inositol phospholipids, and the opening or closing of the potassium, calcium, and chloride ion channels found in certain tissues.

产品编号	M00185
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 cholinergic receptor muscarinic 1 (M1) 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, 200 μg/ml Zeocin (Cat. No. R250-01, 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. Carbacol-induced concentration-dependent stimulation of intracellular calcium mobilization in CHO-K1/M1 cells. The cells were loaded with Calcium-4 prior to being stimulated with agonist carbacol. The intracellular calcium change was normalized and measured by FLIPR. The relative fluorescent units (RFU) were normalized and plotted against the log of the cumulative doses of carbacol (Mean ± SEM, n = 3). The EC₅₀ of carbacol on this cell was 30.92 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 IP-One accumulation upon treatment with carbacol in CHO-K1/M1 cells. d2 acceptor fluorophore-labeled IP-One (Cat. No. 62IPAPEB; Revvity) and intracellular IP-One in CHO-K1/M1 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 carbacol on CHO-K1/M1 cells was 1.39 μM.

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

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CHO-K1/M1 Stable Cell Line

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