| Product Description |
Recombinant CHO-K1 cells stably overexpress human opioid receptor delta 1 (OPRD1) on the surface and contain high levels of G protein Gαi to couple with the receptor in downstream signaling pathways. |
| Applications |
Binding assay, Calcium flux assay, and cAMP accumulation assay. |
| Expressed Gene |
NCBI reference sequence NM_000911; no expressed tags |
| Target Protein |
NP_000902 |
| Host Cell |
CHO-K1 |
| 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. |
| 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) (Cat. No. 21127, Life Technologies), 45% FBS (Cat. No. 10099-141, Life Technologies), 10% DMSO (Cat. No. D2650, Sigma) |
| Complete Growth Medium |
Ham’s F-12K (Kaighn’s), 10% FBS |
| Culture Medium |
Ham’s F-12K (Kaighn’s), 10% FBS, 200 μg/ml Zeocin (Cat. No. R250-01, Life Technologies), 100 μg/ml Hygromycin B (Cat. No. 10687010, Life Technologies) |
| Gene Synonyms |
DOP, DOR, DOR1, OPRD |
| Background |
Opioid receptors and their endogenous peptide ligands play important roles in the reward and reinforcement of drugs such as heroin, cocaine, and alcohol. Opioid receptor family includes three classic receptors, μ, δ, and κ, also known as OP1, OP2 and OP3, respectively. The receptors are Gi/o-coupled GPCRs which will reduce intracellular cAMP levels after activation. δ-opioid receptor modulates many kinase cascades including ERKs, Akts, JNKs, STAT3, P38 involving Src, Ras, Rac, Raf-1, Cdc42, RTKs. In addition, δ-opioid receptor has also been proposed to interact with μ receptors. The observed pharmacological cross-talk may partially arise from agonist cross-reactivity. |
Figure 1. SNC80-induced concentration-dependent stimulation of intracellular calcium mobilization in CHO-K1/OPRD1/Gα15 cells. The cells were loaded with Calcium-4 (Cat. No. R8142; Molecular Devices) prior to stimulation with OPRD1 agonist, SNC80. The intracellular calcium change was measured by FLIPRTETRA. The relative fluorescent units (RFU) were recorded and normalized to plot against the log of the cumulative doses of SNC80 (mean ± SEM, n = 3). The EC50 of SNC80 on CHO-K1/OPRD1/Gα15 cells was 0.75 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 along a sigmoid curve.
Figure 2. Dose dependent stimulation of intracellular cAMP accumulation upon treatment with SNC80 in CHO-K1/OPRD1/Gα15 cells. d2 acceptor fluorophore -labeled cAMP (Cat. No. 62AM4PEC; Revvity) and intracellular cAMP in CHO-K1/OPRD1/Gα15 cells competitively bind with Europium Cryptate-labeled anti-cAMP monoclonal antibody. The FRET signal decreases as the intracellular cAMP concentration rises and was measured by plate reader (Pherastar, BMG). The EC50 of SNC80 on CHO-K1/OPRD1/Gα15 cells was 0.10 μM.
For research use only. Not intended for human or animal clinical trials, therapeutic or diagnostic use.
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