| Product Description |
Recombinant CHO-K1 cells stably overexpress human adrenoceptor alpha 1A (ADRA1A) on the surface and contain high levels of G protein Gαq to couple with the receptor in downstream signaling pathways. |
| Applications |
Binding assay, calcium mobilization assay, IP-One assay. |
| Expressed Gene |
NCBI reference sequence NM_000680; no expressed tags |
| Target Protein |
NP_000671 |
| 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 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, 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, 400 μg/ml Geneticin (Cat. No. 10131-035, Life Technologies) |
| Gene Synonyms |
ADRA1C, ADRA1L1, ALPHA1AAR |
| Background |
The α1-adrenergic receptor (AR) family consists of three closely related gene products (α1A, α1B, and α1D) that mediate the actions of norepinephrine (NE) and epinephrine in sympathetically innervated tissues and brain. α1ARs belong to the G protein-coupled receptor family and consist of single polypeptide chains predicted to have seven transmembrane spanning domains. With similar pharmacological and signaling properties, α1-AR subtypes act through Gq/11 proteins to activate phospholipase C, increase both inositol 1,4,5-trisphosphate production and intracellular Ca2+. Once activated by binding, α1-ARs initiate the cellular pathways leading to the regulation of physiological effects, including blood pressure maintenance, glucose metabolism, renal sodium reabsorption, and cardiac inotropy. |
Figure 1: Epinephrine-induced concentration-dependent stimulation of intracellular calcium mobilization in CHOK1/ADRA1A cells. The cells were loaded with Calcium-4 prior to being stimulated with an ADRA1A receptor agonist, epinephrine. The intracellular calcium change was measured by FLIPR. The relative fluorescent units (RFU) were plotted against the log of the cumulative doses (10-fold dilution) of epinephrine (Mean ± SD, n = 2). The EC50 of epinephrine on this cell was 0.72 nM.
Note:
1. EC50 value is calculated with four parameter logistic equation:
Y=Bottom + (Top-Bottom)/ (1+10^ ((LogEC50-X)*HillSlope))
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.
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