Product Description |
Recombinant CHO-K1 cells stably overexpress human adrenoceptor alpha 2A
(ADRA2A) on the surface and contain high levels of G protein Gαi to couple with
the receptor in downstream signaling pathways. |
Applications |
binding assay,
calcium mobilization assay |
Expressed Gene |
Codon optimized from NM_000681;
no expressed tags |
Target Protein |
NP_000672 |
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). |
Species |
Human |
Stability |
Stable through more than 15 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, 200 μg/ml Zeocin
(Cat. No. R250-01, Life
Technologies), 200
μg/ml Hygromycin B (Cat. No. 10687010,
Invitrogen) |
Gene Synonyms |
ADRA2A; ADRA2;
ADRA2R; ADRAR; ALPHA2AAR; ZNF32 |
Background |
The
adrenoceptor ADRA2A is expressed in the brain, spleen, kidney, aorta, lung,
skeletal muscle, heart and liver. ADRA2A-adrenoceptor knockout mice exhibit
disruption of presynaptic inhibition of noradrenaline release at high
stimulation frequencies. ADRA2A receptor is the principal autoreceptor in the
presynaptic feedback loop regulating noradrenaline release. However, another α2
autoreceptor is also present. |
Figure 1. UK14304-induced concentration-dependent stimulation of intracellular calcium mobilization in CHO-K1/ADRA2A/Gα15 cells. The cells were loaded with Calcium-4 prior to being stimulated with agonist UK14304. 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 UK14304 (Mean ± SEM, n = 3). The EC50 of UK14304 on this cell was 2.68 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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