| 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. |
| Applications |
Calcium mobilization
assay and cAMP accumulation assay. |
| Expressed Gene |
Codon optimized from NM_001052;
no expressed tags |
| Target Protein |
NP_001043 |
| 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, Gibco), 45% FBS (Cat. No. 10099-14,
Gibco), 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 G418 (Cat. No. 10131-035, Gibco), 100 μg/ml
Hygromycin B (Cat. No. 10687010, Invitrogen) |
| Gene Synonyms |
SSTR4 |
| Background |
Somatostatin
acts at many sites to inhibit the release of many hormones and other secretory
proteins. The biologic effects of somatostatin are probably mediated by a
family of G protein-coupled receptors that are expressed in a tissue-specific
manner. SST4 is a member of the superfamily of receptors having seven
transmembrane segments and is expressed in highest levels in fetal and adult
brain and lung. SST4 enhances AMPA receptor-mediated currents in the hippocampus,
and interacts with the SST2 receptor. |
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.
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.
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