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) |
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 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) |
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. |
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.
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