Product Description |
Recombinant CHO-K1/CRF2/Gα15
(cGMP) cells stably overexpress
human corticotropin releasing factor receptor (CRF2) on the surface and contain
high levels of G protein (Gs) to couple with the
receptor in downstream signaling pathways. |
Applications |
Functional assay for
CRF2 receptor (cAMP accumulation assay) |
Expressed Gene |
Codon optimized from NM_001883.5; 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.
# 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% culture medium,
45% FBS (Cat. # A5669701, Gibco), 10%
DMSO (Cat. #D2650, Sigma) |
Complete Growth Medium |
Ham’s F-12 K
(Kaighn’s) (Cat. #21127, Life Technologies), 10% FBS |
Culture Medium |
Ham’s F-12 K (Kaighn’s), 10% FBS, 400 μg/ml G418 (Cat. #10131-035, Gibco),
200 μg/ml Hygromycin B (Cat. #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. # 62AM4PEC, Cisbio) 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 PHERAstar (BMG). The EC50 of Urocortin on CHO-K1/CRF2/Gα15 cells was 0.0865 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 Ratio (665/620) and starts at Bottom and goes to Top along a sigmoid curve.
For research use only. Not intended for human or animal clinical trials,
therapeutic or diagnostic use.
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