安定細胞株製品 » Human Recombinant Corticotropin Releasing Factor Receptor CRF2 Stable Cell Line
CHO-K1/CRF2/Gα15 Stable Cell Line

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.

CHO-K1/CRF2/Gα15 Stable Cell Line

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.
M00208
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Overview
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)

Product Introduction
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 Conditions
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)

Background
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.

Examples
  • CHO-K1/CRF2/Gα15 Stable Cell Line
    • CHO-K1/CRF2/Gα15 Stable Cell Line

    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.