CHO-K1/CXCR4/Gα15 Stable Cell Line

CXCR4 is a receptor for the C-X-C chemokine SDF-1 (Stromal Cell-Derived Factor 1). It is involved in haematopoiesis and cardiac ventricular septum formation, and plays an essential role in vascularization of the gastrointestinal tract, cerebellar development and survival of hippocampal-neuron of central nerve system. CXCR4 also acts as a primary receptor for some HIV-2 isolates and as a co-receptor with CD4 for HIV-1 X4 viruses.

Price	$8500.00
Cat. No.	M00556
Size	2 vials Buy in bulk
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Product Introduction Culture Conditions Bottom Notes

Product Introduction

Product Description	Recombinant CHO-K1 cells stably overexpress human C-X-C motif chemokine receptor 4 (CXCR4) on the surface and contain high levels of G protein Gαi to couple with the receptor in downstream signaling pathways.
Culture Properties	Adherent.
Stability	Stable through more than 16 passages with no 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.

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Culture Conditions

Culture Medium	Ham’s F-12K (Kaighn’s), 10% FBS, 4 μg/ml Puromycin (Cat. No. A11138-03, Life Technologies), 100 μg/ml Hygromycin B (Cat. No. 10687010, Life Technologies)
Complete Growth Medium	Ham’s F-12K (Kaighn’s), 10% FBS
Freeze Medium-DATA	45% Ham’s F-12K (Kaighn’s) (Cat. No. 21127, Life Technologies), 45% FBS (Cat. No. 10099-141, Life Technologies), 10% DMSO (Cat. No. D2650, Sigma)

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Figure 1. CXCL12-induced concentration-dependent stimulation of intracellular calcium mobilization in CHO-K1/Gα15/CXCR4 cells. The cells were loaded with Calcium-4 (Cat. No. R8142; Molecular Devices) prior to stimulation with CXCR4 agonist, CXCL12 (Cat. No. Z02825, Genscript). The intracellular calcium change was measured by FLIPRTETRA. The relative fluorescent units (RFU) were recorded and plotted against the log of the cumulative doses of CXCL12 (mean ± SEM, n = 3). The EC₅₀ of CXCL12 on CHO-K1/Gα15/CXCR4 cells was 0.40 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 along a sigmoid curve.

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Figure 2. Dose dependent stimulation of intracellular cAMP accumulation upon treatment with CXCL12 in CHO-K1/Gα15/CXCR4 cells. d2 acceptor fluorophore-labeled cAMP (Cat. No. 62AM4PEC; Revvity) and intracellular cAMP in CHO-K1/Gα15/CXCR4 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 CXCL12 on CHO-K1/Gα15/CXCR4 cells was 20.76 ng/ml. »

For research use only. Not intended for human or animal clinical trials, therapeutic or diagnostic use.