安定細胞株製品 » Human Recombinant Muscarinic Acetylcholine Receptor M2 Stable Cell Line
CHO-K1/M2/Gα15 Stable Cell Line

Figure 1. Oxotremorine-induced concentration-dependent stimulation of intracellular calcium mobilization in CHO-K1/Gα15/M2 cells. The cells were loaded with Calcium-4 prior to being stimulated with an M2 receptor agonist, Oxotremorine. The intracellular calcium change was measured by FLIPRTETRA. The relative fluorescent units (RFU) were normalized and plotted against the log of the cumulative doses (10-fold dilution) of Oxotremorine (Mean ± SD, n = 2). The EC50 of Oxotremorine on this cell was 63.7 nM.
Notes:
1. EC50 value is calculated with four parameter logistic equation:
Y=Bottom + (Top-Bottom)/(1+10^((LogEC50-X)*HillSlope))
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.

CHO-K1/M2/Gα15 Stable Cell Line

Figure 2. 10 μg of membranes prepared from CHO-K1 cells stably expressing M2 receptors were incubated with indicated concentrations of [3H]N-Methylscopolamine ([3H]NMS) in the absence (total binding) or presence of 1000- fold access unlabeled Atropine (nonspecific binding, NSB). Binding was terminated by rapid filtration. Specific binding was defined by subtracting NSB from total binding. Data were fit to one-site binding equation using a non-linear regression method.

CHO-K1/M2/Gα15 Stable Cell Line

Figure 3. 10 μg of membranes prepared from CHO-K1 cells stably expressing M2 receptors were incubated with indicated concentrations of Atropine in the presence of 0.2 nM [3H]N-Methylscopolamine ([3H]NMS). Binding was terminated by rapid filtration. Data were fit to one-site competition equation using a non-linear regression method.

CHO-K1/M2/Gα15 Stable Cell Line

Muscarinic acetylcholine receptors belong to a superfamily of seven-TM-domain receptors that interact with G-proteins to initiate intracellular responses. Five muscarinic receptor subtypes have been identified and named from M1 to M5. The M2 muscarinic receptor couples to Gi/o to inhibit cAMP production. GenScript co-transfected human M2 with Gα15 in the CHO-K1 which supports high levels of recombinant M2 expression on the cell surface and contains high levels of Gα15 to couple the receptor to the calcium signaling pathway.
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Description

Muscarinic acetylcholine receptors belong to a superfamily of seven-TM-domain receptors that interact with G-proteins to initiate intracellular responses. Five muscarinic receptor subtypes have been identified and named from M1 to M5. The M2 muscarinic receptor couples to Gi/o to inhibit cAMP production. GenScript co-transfected human M2 with Gα15 in the CHO-K1 which supports high levels of recombinant M2 expression on the cell surface and contains high levels of Gα15 to couple the receptor to the calcium signaling pathway.
Synonyms

M2 receptor, m2, acetylcholine receptor, muscarinic 2, Acm2, 7TM receptor, M2 muscarinic acetylcholine receptor, cholinergic receptor, muscarinic 2, cholinergic receptor, muscarinic 2, cardiac, muscarinic acetylcholine receptor M2, muscarinic receptor m2, AChR M2, muscarinic acetylcholine receptor 2, Chrm-2

Overview
Applications Functional assays for M2 receptor

Product Introduction
Storage Liquid nitrogen immediately upon delivery
Species Human

Culture Conditions
Freeze Medium 45% culture medium, 45% FBS (Cat. #10099-141, Gibco), 10% DMSO (Cat. #D2650, Sigma)
Culture Medium Ham’s F-12K (Kaighn’s), 10% FBS, 200 μg/ml Zeocin (Cat. #R250-01, Life Technologies), 100 μg/ml Hygromycin B (Cat. #10687010, Invitrogen)

Examples
  • CHO-K1/M2/Gα15 Stable Cell Line
    • CHO-K1/M2/Gα15 Stable Cell Line

    Figure 1. Oxotremorine-induced concentration-dependent stimulation of intracellular calcium mobilization in CHO-K1/Gα15/M2 cells. The cells were loaded with Calcium-4 prior to being stimulated with an M2 receptor agonist, Oxotremorine. The intracellular calcium change was measured by FLIPRTETRA. The relative fluorescent units (RFU) were normalized and plotted against the log of the cumulative doses (10-fold dilution) of Oxotremorine (Mean ± SD, n = 2). The EC50 of Oxotremorine on this cell was 63.7 nM.
    Notes:
    1. EC50 value is calculated with four parameter logistic equation:
    Y=Bottom + (Top-Bottom)/(1+10^((LogEC50-X)*HillSlope))
    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.

  • CHO-K1/M2/Gα15 Stable Cell Line
    • CHO-K1/M2/Gα15 Stable Cell Line

    Figure 2. 10 μg of membranes prepared from CHO-K1 cells stably expressing M2 receptors were incubated with indicated concentrations of [3H]N-Methylscopolamine ([3H]NMS) in the absence (total binding) or presence of 1000- fold access unlabeled Atropine (nonspecific binding, NSB). Binding was terminated by rapid filtration. Specific binding was defined by subtracting NSB from total binding. Data were fit to one-site binding equation using a non-linear regression method.

  • CHO-K1/M2/Gα15 Stable Cell Line
    • CHO-K1/M2/Gα15 Stable Cell Line

    Figure 3. 10 μg of membranes prepared from CHO-K1 cells stably expressing M2 receptors were incubated with indicated concentrations of Atropine in the presence of 0.2 nM [3H]N-Methylscopolamine ([3H]NMS). Binding was terminated by rapid filtration. Data were fit to one-site competition equation using a non-linear regression method.