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 FLIPR^TETRA. 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

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.

Cat. No.	M00304
Size	2 vials : Quote Buy in bulk
Quantity
Price	¥1,613,997.00


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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

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