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CHO-K1/CXCR4/Gα15 Stable Cell Line

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.

CHO-K1/CXCR4/Gα15 Stable Cell Line

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.

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.
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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.

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)

  • CHO-K1/CXCR4/Gα15 Stable Cell Line
  • CHO-K1/CXCR4/Gα15 Stable Cell Line

    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.

  • CHO-K1/CXCR4/Gα15 Stable Cell Line
  • CHO-K1/CXCR4/Gα15 Stable Cell Line

    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.


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