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目录产品 » 稳定细胞系 » Human Recombinant Muscarinic Acetylcholine Receptor M5 Stable Cell Line
CHO-K1/M5 Stable Cell Line

Figure 1. Carbacol-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 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 0.19 μM.

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.

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.
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产品简介
Product Description Recombinant CHO-K1 cells stably overexpress human cholinergic receptor muscarinic 5 (M5) 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
  • CHO-K1/M5 Stable Cell Line
    • CHO-K1/M5 Stable Cell Line

    Figure 1. Carbacol-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 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 0.19 μM.

    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.


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


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