| Product Description |
Recombinant CHO-K1 cells stably overexpress human calcitonin receptor (CT)
on the surface and contain high levels of G protein Gαq and Gαs to couple with
the receptor in downstream signaling pathways. |
| Applications |
Calcium mobilization
assay, and cAMP accumulation assay. |
| Expressed Gene |
Codon optimized from NM_001742;
no expressed tags |
| Target Protein |
NP_001733 |
| 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,
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), 100
μg/ml Hygromycin B (Cat. No. 10687010,
Invitrogen) |
| Gene Synonyms |
CALCR, CT-R, CTR, CTR1 |
| Background |
Numerous
studies have demonstrated that the calcitonin receptor (CALCR) is a specific
marker of osteoclast differentiation and that calcitonin can inhibit bone
resorption in vitro and in vivo. Mice lacking calcitonin and calcitonin gene–related
peptide (CGRP) have a high bone mass phenotype due to an increase in bone
formation parameters. Expression of calcitonin (CT) and its receptor (CTR)
generate survival, adhesion, pro-inflammatory, and pro-metastatic pathways.
Moreover, data indicate a pivotal role for CT-CTR axis in advanced prostate
cancer PC metastasis and may serve as a potential therapeutic target for
advanced PC. |
Figure 1: Calcitonin-induced concentration-dependent stimulation of intracellular calcium mobilization in CHO-K1/CT/Gα15 cells. The cells were loaded with Calcium-4 (Cat. No. R8142; Molecular Devices) prior to stimulation with CT agonist, calcitonin. 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 calcitonin (mean ± SEM, n = 3). The EC50 of calcitonin on CHO-K1/CT/Gα15 cells was 18.88 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.
Figure 2. Dose dependent stimulation of intracellular cAMP mobilization upon treatment with calcitonin in CHO-K1/CT/Gα15 cells. d2 acceptor fluorophore-labeled cAMP (Cat. No. 62AM4PEC; Revvity) and intracellular cAMP in CHO-K1/CT/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 (Varioskan, Thermo Fisher). The EC50 of calcitonin was 0.27 nM.
For research use only. Not intended for human or animal clinical trials, therapeutic
or diagnostic use.
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