Catalog Products » Stable Cell Lines » Human Recombinant Muscarinic Acetylcholine Receptor M3 Stable Cell Line
CHO-K1/M3 Stable Cell Line

Figure 1. Carbachol-induced concentration-dependent stimulation of intracellular calcium mobilization in CHO-K1/M3 cells. The cells were loaded with Calcium-4 prior to being stimulated with an M3 receptor agonist, Carbachol. 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 Carbachol (Mean ± SD, n = 2). The EC50 of Carbachol on this cell was 12.4 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/M3 Stable Cell Line

Figure 2 10 μg of membranes prepared from CHO-K1 cells stably expressing M3 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/M3 Stable Cell Line

Figure 3 10 μg of membranes prepared from CHO-K1 cells stably expressing M3 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/M3 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 M3 muscarinic receptors are located at many places in the body, e.g. smooth muscles, endocrine, exocrine glands, as well as lungs. They are also found in the CNS, where it induces emesis. They generally cause smooth muscle contraction and increased glandular secretions.
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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 M3 muscarinic receptors are located at many places in the body, e.g. smooth muscles, endocrine, exocrine glands, as well as lungs. They are also found in the CNS, where it induces emesis. They generally cause smooth muscle contraction and increased glandular secretions.

Synonyms

M3 receptor, m3, HM4, acetylcholine receptor, muscarinic 3, cholinergic receptor, muscarinic 3, cholinergic receptor, muscarinic 3, cardiac, muscarinic acetylcholine receptor M3, M3R, Chrm-3, muscarinic acetylcholine receptor 3

Overview
Applications Functional assays for M3 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, 400 μg/ml G418 (Cat. #10131-035, Gibco)

Examples
  • CHO-K1/M3 Stable Cell Line
  • CHO-K1/M3 Stable Cell Line

    Figure 1. Carbachol-induced concentration-dependent stimulation of intracellular calcium mobilization in CHO-K1/M3 cells. The cells were loaded with Calcium-4 prior to being stimulated with an M3 receptor agonist, Carbachol. 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 Carbachol (Mean ± SD, n = 2). The EC50 of Carbachol on this cell was 12.4 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/M3 Stable Cell Line
  • CHO-K1/M3 Stable Cell Line

    Figure 2 10 μg of membranes prepared from CHO-K1 cells stably expressing M3 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/M3 Stable Cell Line
  • CHO-K1/M3 Stable Cell Line

    Figure 3 10 μg of membranes prepared from CHO-K1 cells stably expressing M3 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.


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