Catalog Products » Stable Cell Lines » Human Recombinant Muscarinic Acetylcholine Receptor M5 Stable Cell Line
CHO-K1/M5 Stable Cell Line

Figure 2. 10 μg of membranes prepared from CHO-K1 cells stably expressing M5 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/M5 Stable Cell Line

Figure 3. 10 μg of membranes prepared from CHO-K1 cells stably expressing M5 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/M5 Stable Cell Line

Figure 1: Carbachol-induced concentration-dependent stimulation of intracellular calcium mobilization in CHO-K1/M5 cells. The cells were loaded with Calcium-4 prior to being stimulated with an M5 receptor agonist, carbachol. The intracellular calcium change was measured by FLIPRTETRA. The relative fluorescent units (RFU) were plotted against the log of the cumulative doses of carbachol (Mean ± SD, n = 2). The EC50 of carbachol on this cell was 188.4 nM.
Note:
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 and starts at Bottom and goes to Top with a sigmoid shape.

CHO-K1/M5 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, named M1 through M5. Receptors of the M5 receptor subtype couple through the Gq/11 class of G-proteins and activate the phospholipase C pathway. Activation of this pathway in turn leads to increases in free intracellular calcium levels as inositol triphosphate mediates release of calcium from the endoplasmic reticulum. RT-PCR reveals that M5 mRNA is quite uniformly expressed in brain. However, there is little data regarding the expression and function of the M5 receptor in peripheral tissues. Currently, it is clear that the M5 receptor, due to the high likelihood that its distribution is restricted to the CNS, probably plays a discrete role in dopaminergic transmission. Although the identification of M5 expression in salivary glands and iris-ciliary muscle suggests a broader role, the data on this is sparse and requires extensive confirmation.
M00186
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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, named M1 through M5. Receptors of the M5 receptor subtype couple through the Gq/11 class of G-proteins and activate the phospholipase C pathway. Activation of this pathway in turn leads to increases in free intracellular calcium levels as inositol triphosphate mediates release of calcium from the endoplasmic reticulum. RT-PCR reveals that M5 mRNA is quite uniformly expressed in brain. However, there is little data regarding the expression and function of the M5 receptor in peripheral tissues. Currently, it is clear that the M5 receptor, due to the high likelihood that its distribution is restricted to the CNS, probably plays a discrete role in dopaminergic transmission. Although the identification of M5 expression in salivary glands and iris-ciliary muscle suggests a broader role, the data on this is sparse and requires extensive confirmation.

Synonyms

M5 receptor, m5, acetylcholine receptor, muscarinic 5, cholinergic receptor, muscarinic 5, muscarinic acetylcholine receptor M5, muscarinic acetylcholine receptor 5, M5R

Overview
Applications Functional assay for M5 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)

Examples
  • CHO-K1/M5 Stable Cell Line
  • CHO-K1/M5 Stable Cell Line

    Figure 2. 10 μg of membranes prepared from CHO-K1 cells stably expressing M5 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/M5 Stable Cell Line
  • CHO-K1/M5 Stable Cell Line

    Figure 3. 10 μg of membranes prepared from CHO-K1 cells stably expressing M5 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/M5 Stable Cell Line
  • CHO-K1/M5 Stable Cell Line

    Figure 1: Carbachol-induced concentration-dependent stimulation of intracellular calcium mobilization in CHO-K1/M5 cells. The cells were loaded with Calcium-4 prior to being stimulated with an M5 receptor agonist, carbachol. The intracellular calcium change was measured by FLIPRTETRA. The relative fluorescent units (RFU) were plotted against the log of the cumulative doses of carbachol (Mean ± SD, n = 2). The EC50 of carbachol on this cell was 188.4 nM.
    Note:
    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 and starts at Bottom and goes to Top with a sigmoid shape.


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