| Product Description |
Recombinant 1321N1 cells stably overexpress human pyrimidinergic receptor P2Y6 (P2Y6) on the surface and contain high levels of G protein Gαq to couple with the receptor in downstream signaling pathways. |
| Applications |
Calcium mobilization assay, IP-One assay. |
| Expressed Gene |
NCBI reference sequence NM_176798; no expressed tags |
| Target Protein |
NP_789768 |
| Host Cell |
1321N1 |
| 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% DMEM (Cat. No. 10569, Life Technologies), 45% FBS (Cat. No. 10099-141, Life Technologies), 10% DMSO (Cat. No. D2650, Sigma) |
| Complete Growth Medium |
DMEM, 10% FBS |
| Culture Medium |
DMEM, 10% FBS, 100 μg/ml Geneticin (Cat. No. 10131-035, Life Technologies) |
| Gene Synonyms |
P2RY6 |
| Background |
P2Y receptors are one of three families of extracellular receptors for purine and pyrimidine nucleotides involved in purinergic signaling. In heterologous expression systems, recombinant P2Y1, P2Y2, P2Y4, P2Y6 receptors couple to G proteins (mainly Gq) which, in turn, activate intracellular signaling cascades - mainly the phospholipase Cβ (PLCβ) isoform to mobilize intracellular Ca2+ ions. RT-PCR and Northern Blotting studies show a high expression of this receptor in spleen and placenta, followed by kidney, adipose, bone, lung, heart, brain and skeletal muscle. In mouse bone marrow-derived mast cells (mBMMCs) and in the human MC line LAD2, P2Y6 receptors cooperate with CysLT1R to promote cell survival and chemokine generation by a pathway involving reciprocal ligand-mediated cross-talk. In human, rat and mouse, P2Y6 expression is increased by the stress associated with intestinal inflammation, leading to the release of CXC chemokine ligand 8. In rabbit, the receptor functions by increasing osteoclast survival through the NFκB pathway |
Figure 1: UDP-induced concentration-dependent stimulation of intracellular calcium mobilization in 1321N1/P2Y6 cells. The cells were loaded with Calcium-4 prior to stimulation with a P2Y6 receptor agonist, UDP. The intracellular calcium change was measured by FLIPRTETRA. The relative fluorescent units (RFU) were normalized and plotted against the log of the cumulative doses (5-fold dilution) of UDP (Mean ± SEM, n = 3). The EC50 of UDP on 1321N1/P2Y6 cells was 75.49 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 % stimulation of RFU and starts at Bottom and goes to Top with a sigmoid shape.
For research use only. Not intended for human or animal clinical trials, therapeutic or diagnostic use.
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