安定細胞株製品 » Human Recombinant μ-Opioid Receptor OPRM1 Stable Cell Line
CHO-K1/OPRM1/Gα15 Stable Cell Line

Figure 1. DAMGO-induced concentration-dependent stimulation of intracellular calcium mobilization in CHO-K1/OPRM1/Gα15 cells. The cells were loaded with Calcium-4 prior to being stimulated with agonist DAMGO. The intracellular calcium change was measured by FLIPRTETRA. The relative fluorescent units (RFU) were normalized and plotted against the log of the cumulative doses of DAMGO (Mean ± SD, n = 2). The EC50 of DAMGO on this cell was 4.2 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/OPRM1/Gα15 Stable Cell Line

CHO-K1/OPRM1/Gα15 Stable Cell Line

CHO-K1/OPRM1/Gα15 Stable Cell Line

The µ-opioid receptor (oprm1) is the principal site of action in the brain by which morphine, other opiate drugs of abuse, and endogenous opioid peptides effect analgesia and alter mood. Opioid receptors belong to the rhodopsin family of G protein-coupled receptors (GPCRs). The three types of opioid receptors (μ, δ, and κ) have been shown to associate with each other in a homotypic or heterotypic fashion when expressed in heterologous cells. A member of the seven-transmembrane domain (TM) G protein-coupled receptor (GPCR) superfamily, the µ-opioid receptor modulates ion channels and second messenger effectors in an opioid agonist-dependent fashion that is reversible by the classic opiate antagonist naloxone.
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Overview
Product Description Recombinant CHO-K1 cells stably co-overexpress human opioid receptor mu 1 (OPRM1) and G15 alpha subunit. Naturally, OPRM1 is an inhibitory G-protein coupled receptor that activates the Gi alpha subunit, inhibiting adenylate cyclase activity, lowering cAMP levels naturally. But when Gα15 and OPRM1 are co-expressed, the cells can also conduct Gq-coupled signal transduction and mobilize intracellular calcium following certain ligand binding. This cell line is recommended for screening and functional validating agonists of OPRM1.
Applications Screening and functional validation of OPRM1 agonists (e.g. Calcium flux assay, cAMP assay)

Product Introduction
Expressed Gene Codon Optimized from NM_000914; no expressed tags
Target Protein NP_000905
Host Cell CHO-K1/Gα15
Quantity 2 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.
Mycoplasma Status Negative. The mycoplasma test was performed with MycoAlert™ PLUS Mycoplasma Detection Kit (Cat. No.LT07-318, Lonza).
Stability Stable through more than 16 passages with no significant changes in assay performance or expression profile.

Culture Conditions
Culture Properties Adherent
Freeze Medium 45% Ham’s F-12K (Kaighn's) Medium (Cat. No. 21127-022, Gibco), 45% FBS (Cat. No. 10099-141, Gibco), 10% (V/V) DMSO (Cat. No. D2650, Sigma)
Complete Growth Medium Ham's F-12K (Kaighn's) Medium (Cat. No. 21127-022, Gibco), 10% FBS (Cat. No. 10099-141, Gibco)
Culture Medium Ham’s F-12K (Kaighn’s) Meidum (Cat. No. 21127-022, Gibco), 10% FBS (Cat. No. 10099-141, Gibco), 200 μg/ml Zeocin (Cat. No. R250-01, Life Technologies), 100 μg/ml Hygromycin B (Cat. No. 10687010, Invitrogen)

Background
Gene Synonyms MOR-1, MOR1, KIAA0403, OPRM
Background Opioid receptor mu 1 (oprm1), also known as µ-opioid receptor, is the principal site of action in the brain by which morphine, other opiate drugs of abuse, and endogenous opioid peptides that affect analgesia and alter mood. Opioid receptors belong to the rhodopsin family of G protein-coupled receptors (GPCRs). The three types of opioid receptors (μ, δ, and κ) have been shown to associate with each other in a homotypic or heterotypic fashion when expressed in heterologous cells. A member of the seven-transmembrane domain (TM) G protein-coupled receptor (GPCR) superfamily, the µ-opioid receptor modulates ion channels and second messenger effectors in an opioid agonist-dependent fashion that is reversible by the classic opiate antagonist naloxone.

Examples
  • CHO-K1/OPRM1/Gα15 Stable Cell Line
  • CHO-K1/OPRM1/Gα15 Stable Cell Line

    Figure 1. DAMGO-induced concentration-dependent stimulation of intracellular calcium mobilization in CHO-K1/OPRM1/Gα15 cells. The cells were loaded with Calcium-4 prior to being stimulated with agonist DAMGO. The intracellular calcium change was measured by FLIPRTETRA. The relative fluorescent units (RFU) were normalized and plotted against the log of the cumulative doses of DAMGO (Mean ± SD, n = 2). The EC50 of DAMGO on this cell was 4.2 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/OPRM1/Gα15 Stable Cell Line
  • CHO-K1/OPRM1/Gα15 Stable Cell Line

  • CHO-K1/OPRM1/Gα15 Stable Cell Line
  • CHO-K1/OPRM1/Gα15 Stable Cell Line