| Product Description |
Recombinant CHO-K1 cells stably overexpress human adrenoceptor alpha 1B (ADRA1B) on the surface and contain high levels of G protein Gαq to couple with the receptor in downstream signaling pathways. |
| Applications |
Binding assay, Calcium flux assay, IP-One assay. |
| Expressed Gene |
NCBI reference sequence NM_000679; no expressed tags |
| Target Protein |
NP_000670 |
| 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 |
ADRA1, ALPHA1BAR |
| 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 CHO-K1/ADRA1B cells. The cells were loaded with Calcium-4 prior to being stimulated with agonist Epinephrine. 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 Epinephrine (Mean ± SEM, n = 3). The EC50 of Epinephrine on this cell was 17.55 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 RFU and starts at Bottom and goes to Top with a sigmoid shape.
Figure 2. Dose dependent stimulation of intracellular IP-One accumulation upon treatment with epinephrine in CHO-K1/ADRA1B cells. d2 acceptor fluorophore-labeled IP-One (Cat. No. 62IPAPEB; Revvity) and intracellular IP-One in CHO-K1/ADRA1B cells competitively bind with Europium Cryptate-labeled anti-IP-One antibody. The FRET signal decreases as the intracellular IP-One concentration rises and was measured by plate reader (Pherastar, BMG). The EC50 of epinephrine on CHO-K1/ADRA1B cells was 0.11 μM.
For research use only. Not intended for human or animal clinical trials, therapeutic or diagnostic use.
如您需要获取细胞系产品的说明书，请点击“Manual”，完善信息后点击“提交”。
我们会第一时间联系回复您。注意：中国地区购买的产品，说明书需要以中文站为准。
您也可拨打400-0258686转5810，或发邮件至product@genscript.com.cn联系我们。
更多产品文件类型 >>
