Product Description |
Recombinant CHO-K1 cells stably
overexpress mas-related G-protein coupled receptor member X2 (MRGPRX2). MRGPRX2
couples to both Gi and Gq alpha subunits in mast cells naturally, so that the
PLC signaling can be triggered within the cells by agonist binding. This cell
line is recommended for agonist screening and functional validations with
calcium flux assay. |
Applications |
Agonist screening and functional validations of MRGPRX2. |
Expressed Gene |
Codon Optimized from NM_054030; no expressed tags |
Target Protein |
NP_473371 |
Host Cell |
CHO-K1 |
Quantity |
2 vials of frozen cells (>1X106 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 15 passages with no
significant changes in assay performance or expression profile. |
Culture Properties |
Adherent |
Freeze Medium |
45% culture medium, 45% FBS (Cat. No. 10099-141,
Gibco), 10% DMSO (Cat. No. D2650, Sigma) |
Complete Growth Medium |
Ham’s F-12K (Kaighn’s) Medium (Cat. No. 21127-022, Gibco),
10% FBS (Cat. No. 10099-141, Gibco) |
Culture Medium |
Ham’s F-12K (Kaighn’s) Medium (Cat. No. 21127-022, Gibco),
10% FBS (Cat. No. 10099-141, Gibco), 200 μg/ml Zeocin (Cat. No. R250-01, Life
Technologies) |
Gene Synonyms |
MRGPRX2; MRGX2 |
Background |
MRGPRX2 (also known as MRGX2) is a Gq and Gi-coupled GPCR that binds to several peptides,
including the neuropeptide cortistatin-14 and the proadrenomedullin N-terminal
peptide (PAMP). It is probably involved in the function of nociceptive neurons
and regulation of nociceptor function and/or development, including the
sensation or modulation of pain. Dorsal root ganglia and adrenal chromaffin
cells express MRGPRX2,
and binding of these ligands to MRGPRX2 is thought to play a role in nociception and catecholamine release from
the adrenal glands. |
Figure 1. Cortistatin 14-induced concentration-dependent stimulation of intracellular calcium mobilization in CHO-K1/MRGPRX2 cells. The cells were loaded with Calcium-4 prior to being stimulated with agonist Cortistatin 14. The intracellular calcium change was measured by FLIPR. The relative fluorescent units (RFU) were normalized and plotted against the log of the cumulative doses (5-fold dilution) of Cortistatin 14 (Mean ± SD, n = 2). The EC50 of Cortistatin 14 on this cell was 0.64 µM.
Notes:
1. EC50 value is calculated with four parameter logistic equation:
Y=Bottom + (Top-Bottom)/(1+10^((LogEC50-X)*HillSlope))
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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