安定細胞株製品 » Human Recombinant PAR2 Proteinase-activated Receptor Stable Cell Line
CHO-K1/PAR2/Gα15 Stable Cell Line

Figure 1. Trypsin-induced concentration-dependent stimulation of intracellular calcium mobilization in CHO-K1/PAR2/Gα15 cells. The cells were loaded with Calcium-4 prior to being stimulated with agonist glucagon. The intracellular calcium change was normalized and measured by FLIPR. The relative fluorescent units (RFU) were normalized and plotted against the log of the cumulative doses of trypsin (Mean ± SEM, n = 3). The EC50 of trypsin on this cell was 4.42 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 RFU and starts at Bottom and goes to Top with a sigmoid shape.

CHO-K1/PAR2/Gα15 Stable Cell Line

Recombinant CHO-K1 cells stably overexpress human protease-activated receptor 2 (PAR2) on the surface and contain high levels of G protein Gαq to couple with the receptor in downstream signaling pathways.
M00446
¥1,613,997.00

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Overview
Product Description Recombinant CHO-K1 cells stably overexpress human protease-activated receptor 2 (PAR2) on the surface and contain high levels of G protein Gαq to couple with the receptor in downstream signaling pathways.
Applications Calcium flux assay.

Product Introduction
Expressed Gene NCBI reference sequence NM_005242; no expressed tags
Target Protein NP_005233
Host Cell CHO-K1
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 Conditions
Culture Properties Adherent
Freeze Medium 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)
Complete Growth Medium Ham’s F-12K (Kaighn’s), 10% FBS
Culture Medium Ham’s F-12K (Kaighn’s), 10% FBS, 200 μg/ml Zeocin (Cat. No. R250-01, Life Technologies), 100 μg/ml Hygromycin B (Cat. No. 10687010, Life Technologies)

Background
Gene Synonyms GPR11, F2RL1
Background Protease-activated receptors (PAR) are a subfamily of G-protein coupled, seven-transmembrane domain receptors, which are cleaved within the aminoterminal exodomain by certain serine proteases at a specific peptide bond. Trypsin and mast cell tryptase, and more recently, the activated coagulation factors VIIa and Xa, have been identified as serine proteases able to activate mammalian PAR-2. As already indicated, PAR-2 is believed to be involved in inflammation. This role for PAR-2 implies that elastase and cathepsin G would paradoxically display an anti-inflammatory property by disarming PAR-2.

Examples
  • CHO-K1/PAR2/Gα15 Stable Cell Line
    • CHO-K1/PAR2/Gα15 Stable Cell Line

    Figure 1. Trypsin-induced concentration-dependent stimulation of intracellular calcium mobilization in CHO-K1/PAR2/Gα15 cells. The cells were loaded with Calcium-4 prior to being stimulated with agonist glucagon. The intracellular calcium change was normalized and measured by FLIPR. The relative fluorescent units (RFU) were normalized and plotted against the log of the cumulative doses of trypsin (Mean ± SEM, n = 3). The EC50 of trypsin on this cell was 4.42 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 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.