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| $8500.00 | |
| M00933 | |
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| Product Description | Recombinant CHO-K1 cells stably overexpress human C-C motif chemokine receptor 9 (CCR9) on the surface and contain high levels of G protein Gαi/o to couple with the receptor in downstream signaling pathways. |
| Culture Properties | Adherent |
| Stability | Stable through more than 20 passages without 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, 100 μg/ml Hygromycin B (Cat. No. 10687010, Life Technologies), 4 μg/ml Puromycin (Cat. No. A11138-03, Life Technologies) |
| Complete Growth Medium | Ham’s F-12K (Kaighn’s) (Cat. No. 21127-022, Life Technologies), 10% FBS (Cat. No. 10099-141, Life Technologies) |
| Freeze Medium-DATA | 90% complete growth medium, 10% (V/V) DMSO (Cat. No. D2650, Sigma) |
Figure 2. Dose dependent stimulation of intracellular cAMP accumulation upon treatment with CCL25 in CHO-K1/Gα15/CCR9 cells. d2 acceptor fluorophore-labeled cAMP (Cat. No. 62AM4PEC; Revvity) and intracellular cAMP in CHO-K1/Gα15/CCR9 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 (Pherastar, BMG). The EC50 of CCL25 on CHO-K1/Gα15/CCR9 cells was 5.64 μg/ml. »
Figure 3. FACS analysis of cell surface expression of CCR9 on CHO-K1/Gα15/CCR9 cells. The CHO-K1/Gα15/CCR9 cells (Red) and the negative control CHO-K1/Gα15 cells (Green) were probed using Alexa Fluor® 647 conjugated anti-human CCR9 (Cat. No. 358912, Biolegend). »
Figure 1. CCL25-induced concentration-dependent stimulation of intracellular calcium mobilization in CHO-K1/Gα15/CCR9 cells. The cells were loaded with Calcium-4 (Cat. No. R8142; Molecular Devices) prior to being stimulated with agonist CCL25 (Cat. No. Z02847; Genscript). The intracellular calcium change was measured by FLIPR. The relative fluorescent units (∆RFU) were plotted against the log of the cumulative doses of CCL25 (Mean ± SEM, n = 3). The EC50 of CCL25 on this cell was 1.27 µg/ml.
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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For research use only. Not intended for human or animal clinical trials, therapeutic or diagnostic use.
