安定細胞株製品 » Human Recombinant Muscarinic Acetylcholine Receptor M4 Stable Cell Line
CHO-K1/M4/Gα15 Stable Cell Line

Figure 1. Oxotremorine M-induced concentration-dependent stimulation of intracellular calcium mobilization in CHO-K1/M4/Gα15 cells. The cells were loaded with Calcium-4 prior to be stimulated with an M4 receptor agonist, oxotremorine M. The intracellular calcium change was measured by FLIPRTETRA. The relative fluorescent units (RFU) were plotted against the log of the cumulative doses (5-fold dilution) of oxotremorine (Mean ± SD, n = 2). The EC50 of oxotremorine M on this cell was 88.7 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.
2. Signal to background Ratio (S/B) = Top/Bottom

CHO-K1/M4/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 M4 muscarinic receptor couples to Gi/o to inhibit cAMP production. GenScript co-transfected human M4 with Gα15 in the CHO-K1 which supports high levels of recombinant M4 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 M4 muscarinic receptor couples to Gi/o to inhibit cAMP production. GenScript co-transfected human M4 with Gα15 in the CHO-K1 which supports high levels of recombinant M4 expression on the cell surface and contains high levels of Gα15 to couple the receptor to the calcium signaling pathway.
Synonyms

M4 receptor, m4, HM3, acetylcholine receptor, muscarinic 4, cholinergic receptor, muscarin 4, muscarinic acetylcholine receptor M4, Chrm-4, muscarinic acetylcholine receptor 4

Overview
Applications Functional assays for M4 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/M4/Gα15 Stable Cell Line
    • CHO-K1/M4/Gα15 Stable Cell Line

    Figure 1. Oxotremorine M-induced concentration-dependent stimulation of intracellular calcium mobilization in CHO-K1/M4/Gα15 cells. The cells were loaded with Calcium-4 prior to be stimulated with an M4 receptor agonist, oxotremorine M. The intracellular calcium change was measured by FLIPRTETRA. The relative fluorescent units (RFU) were plotted against the log of the cumulative doses (5-fold dilution) of oxotremorine (Mean ± SD, n = 2). The EC50 of oxotremorine M on this cell was 88.7 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.
    2. Signal to background Ratio (S/B) = Top/Bottom