| Product Description |
Recombinant CHO-K1 cells stably overexpress human apelin receptor (AGTRL1) on the surface and contain high levels of G protein Gαi to couple with the receptor in downstream signaling pathways. |
| Applications |
cAMP accumulation assay, calcium flux assay. |
| Expressed Gene |
NCBI reference sequence NM_005161; no expressed tags |
| Target Protein |
NP_005152 |
| 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, 400 μg/ml Geneticin (Cat. No. 10131-035, Life Technologies), 100 μg/ml Hygromycin B (Cat. No. 10687010, Life Technologies) |
| Gene Synonyms |
APLNR, APJ, APJR, HG11 |
| Background |
The apelin receptor AGTRL1 is Gi/Go-coupled GPCRs expressed in the heart, coronary artery, aorta, internal mammary artery, pulmonary artery and saphenous vein, it also expressed in lung, kidney and adrenal gland. In AGTRL1 receptor knockout mice baseline blood pressure is not changed compared to wild-type animals. Vasoconstrictor responses to angiotensin II however are significantly more pronounced in the knockout animals. |
Figure 2. Apelin 13-induced concentration-dependent stimulation of intracellular calcium mobilization in CHO-K1/ AGTRL1/Gα15 cells. The cells were loaded with Calcium-4 (Cat. No. R8142, Molecular Devices) prior to stimulation with AGTRL1 receptor agonist, apelin 13. The intracellular calcium change was measured by FLIPRTETRA. The relative fluorescent units (∆RFU) were normalized and plotted against the log of the cumulative doses (5-fold dilution) of apelin 13 (Mean ± SEM, n = 3). The EC50 of apelin 13 on this cell was 0.18 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.
Figure 1. Dose dependent stimulation of intracellular cAMP accumulation upon treatment with Apelin-13 in CHO-K1/AGTRL1/Gα15 cells. d2 acceptor fluorophore -labeled cAMP (Cat. No. 62AM4PEC; Revvity) and intracellular cAMP in CHO-K1/AGTRL1/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 Apelin-13 on CHO-K1/AGTRL1/Gα15 cells was 9.11 nM.
For research use only. Not intended for human or animal clinical trials, therapeutic or diagnostic use.
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