| Product Description |
Human Recombinant CHO-K1/CCKB Stable Cell Line
stably overexpress the Cholecystokinin Receptor gene (CCKB). This stable cell line product is designed for measuring
the In Vitro function of CCKB. |
| Applications |
Functional assay for CCKB receptor (Calcium Flux)
|
| Expressed Gene |
Codon optimized from
NM_176875; no expressed tags
|
| Target Protein |
NP_795344.1
|
| 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 without significant changes in
assay performance or expression profile.
|
| Culture Properties |
Adherent
|
| Freeze Medium |
Ham’s F-12K (Kaighn’s) (Cat. No. 21127, Gibco), 45% FBS (Cat. No. A5669701,
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, 200 μg/ml Zeocin (Cat. No. R250-01,
Life Technologies)
|
| Gene Synonyms |
CCK-2R; CCK-B; CCK2R; GASR;
CCKBR
|
| Background |
Cholecystokinins (CCK) are derived from preprocholecystokinin.
They are widely expressed in gastrointestinal tissues and in the CNS. There are
two cholecystokinin receptors, referred to as A and B (CCKAR and CCKBR). CCKBR
can be activated by the smaller CCK-4. In the CNS, CCKBR has been implicated in
anxiety, depression, schizophrenia, depression, and opioid analgesia. In the
stomach, CCK-2 mediates gastrin-stimulated gastric acid secretion. |
Figure 1: CCK8-induced concentration-dependent stimulation of intracellular calcium mobilization in CHO-K1/CCKB cells.
The cells were loaded with Calcium-4 (Cat. No. R8141; Molecular Devices) prior to stimulation with CCKB agonist, CCK8. 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 CCK8 (mean ± SD, n = 3). The EC50 of CCK8 on CHO-K1/CCKB cells was 0.945 nM.
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.
For research use only. Not intended for human or animal
clinical trials, therapeutic or diagnostic use.
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