| Product Description |
Recombinant CHO-K1 cells stably overexpress human corticotropin releasing hormone receptor 2 (CRF2) on the surface and contain high levels of G protein Gαs to couple with the receptor in downstream signaling pathways. |
| Applications |
cAMP accumulation assay. |
| Expressed Gene |
NCBI reference sequence NM_001883.2; no expressed tags |
| Target Protein |
NP_001874.2 |
| 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 Properties |
Adherent |
| Freeze Medium |
45% Ham’s F-12K (Kaighn’s) (Cat. No. 21127, Life Technologies), 45% FBS (Cat. No. 10099-141, Gibco), 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, 400 μg/ml Geneticin (Cat. No. 10131-035, Life Technologies), 200 μg/ml Hygromycin B (Cat. No. 10687010, Invitrogen) |
| Gene Synonyms |
CRF-RB; CRHR2; CRFR2; HM-CRF |
| Background |
The corticotropin-releasing factor receptor 2 CRF2 is Gs-coupled GPCRs expressed in the brain, blood vessels and intestine that bind to corticotropin-releasing factor (CRF). The CRF is a 41-amino acid peptide that plays an important role in the integration of autonomic, neuroendocrine, and behavioral responses to stress. GenScript generated a clonal human CRF2-expressing cell line co-expressing Gα15 in the CHO-K1 host cells. |
Figure 1. Dose dependent stimulation of intracellular cAMP accumulation upon treatment with urocortin in CHO-K1/CRF2/Gα15 cells. d2 acceptor fluorophore -labeled cAMP (Cat. No. 62AM4PEC; Revvity) and intracellular cAMP in CHO-K1/CRF2/Gα15 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 urocortin on CHO-K1/CRF2/Gα15 cells was 0.086 nM.
For research use only. Not intended for human or animal clinical trials, therapeutic or diagnostic use.
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