| Product Description |
Recombinant CHO-K1 cells stably overexpress human somatostatin receptor 5 (SST5) on the surface and contain high levels of G protein Gαi to couple with the receptor in downstream signaling pathways. |
| Applications |
Calcium flux assay, IP-One assay and cAMP accumulation assay. |
| Expressed Gene |
NCBI reference sequence NM_001053; no expressed tags |
| Target Protein |
NP_001044 |
| 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 Technologie s), 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 |
SS-5-R, SST5R |
| Background |
Somatostatin receptors (SSTRs), a family of seven transmembrane (TM) domain G-protein-coupled receptors having five distinct subtypes (termed SSTR1–5), are activated by somatostatin secreted from the nerve and endocrine cells. SSTRs are widely expressed in many tissues, frequently as multiple subtypes that coexist in the same cell. With expressions in a tissue-specific manner, SSTRs are involved in the regulation of secretion of insulin, glucagon and growth hormone as well as cell growth induced by neuronal excitation in both the central and peripheral nervous systems. The five receptors share common signaling pathways such as the inhibition of adenylyl cyclase, activation of phosphotyrosine phosphatase (PTP), and modulation of mitogen-activated protein kinase (MAPK) through G-protein-dependent mechanisms. Aberrant expression of somatostatin receptors is known to be involved in a large number of human tumors. |
Figure 1. Somatostatin-14-induced concentration-dependent stimulation of intracellular calcium mobilization in CHO-K1/SST5/Gα15 cells. The cells were loaded with Calcium-4 (Cat. No. R8142; Molecular Devices) prior to stimulation with SST5 agonist, somatostatin-14. 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 somatostatin-14 (mean ± SEM, n = 3). The EC50 of somatostatin-14 on CHO-K1/SST5/Gα15 cells was 0.35 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 % stimulation of RFU and starts at Bottom and goes to Top along a sigmoid curve.
Figure 2. Dose dependent stimulation of intracellular IP-One accumulation upon treatment with somatostatin-14 in CHO-K1/SST5/Gα15 cells. d2 acceptor fluorophore-labeled IP-One (Cat. No. 62IPAPEB; Revvity) and intracellular IP-One in CHO-K1/SST5/Gα15 cells competitively bind with Europium Cryptate-labeled anti-IP-One antibody. The FRET signal decreases as the intracellular IP-One concentration rises and was measured by plate reader (Pherastar, BMG). The EC50 of somatostatin-14 on CHO-K1/SST5/Gα15 cells was 0.19 μM.
For research use only. Not intended for human or animal clinical trials, therapeutic or diagnostic use.
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