Product Description | Recombinant CHO-K1 cells stably overexpress human somatostatin receptor 4 (SST4) on the surface and contain high levels of G protein Gαi to couple with the receptor in downstream signaling pathways. |
Culture Properties | Adherent. |
Stability | Stable through more than 16 passages with no significant changes in assay performance or expression profile. |
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. |
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) |
Complete Growth Medium | Ham’s F-12K (Kaighn’s), 10% FBS |
Freeze Medium-DATA | 45% Ham’s F-12K (Kaighn’s) (Cat. No. 21127, Life Technologies), 45% FBS (Cat. No. 10099-14, Life Technologies), 10% DMSO (Cat. No. D2650, Sigma). |
Figure 1. Somatostatin-14-induced concentration-dependent stimulation of intracellular calcium mobilization in CHO-K1/SST4/Gα15 cells. The cells were loaded with Calcium-4 prior to being stimulated with agonist somatostatin-14. The intracellular calcium change was measured by FLIPR. The relative fluorescent units (RFU) were normalized and plotted against the log of the cumulative doses of Somatostatin-14 (Mean ± SEM, n = 3). The EC50 of Somatostatin-14 on this cell was 0.98 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.
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Figure 2. Dose dependent changing of intracellular cAMP upon treatment with Somatostain-14 in CHO-K1/SST4/Gα15 cells. d2 acceptor fluorophore-labeled cAMP and intracellular cAMP in CHO-K1/SST4/Gα15 cells competitively bind with Europium-labeled anti-cAMP monoclonal antibody. The FRET signal decreases as the intracellular cAMP concentration rises and was measured by PHERAstar FSX (BMG). The EC50 of Somatostatin-14 on CHO-K1/SST4/Gα15 cells was 5.26 nM. »
For research use only. Not intended for human or animal clinical trials, therapeutic or diagnostic use.