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Calcium flux in β2-expressing Chem-2 cell line induced by Formoterol. β2-expressing Chem-2 cells (Chem2-β2) and Wild-Type Chem-2 cells (Chem2-WT) wer ...
ChemiScreen™ β2 Adrenoceptor Calcium-Optimized FLIPR Cell Lines
Description:
ChemiScreen™ β2 Adrenoceptor Calcium-Optimized FLIPR Cell Lines
Trade Name:
ChemiScreen
Product Overview:
Full-length human ADRB2 cDNA encoding the β2-adrenoceptor
Background Information:
The endogenous catecholamines epinephrine and norepinephrine have profound effects on smooth muscle activity, cardiac function, carbohydrate and fat metabolism, hormone secretion, neurotransmitter release, and central nervous system actions. These activities are mediated by GPCRs belonging to two subfamilies, the α- and β-adrenergic receptors (Bylund et al., 1994). The β-adrenergic receptors, primarily the β2 subtype, mediate relaxation of smooth muscle in many tissues, and β 2-selective agonists are the preferred drugs for stimulating bronchodilation in the treatment of asthma and chronic obstructive pulmonary disease (Sears and Lotvall, 2005). Activation of the β-adrenergic receptors, primarily the β1 subtype and to a lesser extent the β2 subtype, acutely increases heart rate, cardiac output, and cardiac automaticity, and chronically increases cardiac myocyte apoptosis. As a result, β-adrenergic receptor antagonists (β blockers) are effective in the treatment of congestive heart failure and arrhythmia (Feldman et al., 2005). Chemicon's cloned human β2-expressing cell line is made in the Chem-2 host, which supports high levels of recombinant β2 expression on the cell surface and contains high levels of the promiscuous G protein Gα16 to couple the receptor to the calcium signaling pathway. Thus, the cell line is an ideal tool for screening for agonists and antagonists of β2.
Application Notes:
Calcium flux assay, ligand binding assays
Species Reactivity:
Human
Usage Statement:
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
Cell Line Type:
GPCR Cell Lines
Concentration:
2 x 106 cells/mL
Host Cells:
Chem-2, a suspension cell line with high levels of the promiscuous G protein Gα16.
Packaging:
1 mL per vial
Materials Required but Not Delivered:
RPMI-1640/10% heat inactivated fetal bovine serum/0.4 mg/mL Geneticin (G418) /100 U/mL each penicillin and streptomycin
Cell Type:
Chem-2
Gene Symbol:
- ADRBK2
- BARK2
- GRK3
- Beta-ARK-2
- EC 2.7.11.15
Presentation:
Cells are frozen at 2 x 106 cells/mL in Cell Culture Freezing Media with DMSO (Chemicon catalog number S-002-10F). Cell line tests negative for mycoplasma.
Quality Assurance:
SPECIFICATIONS: EC50 for calcium mobilization by Formoterol ~ 6 nM
Protein or Enzyme Type:
GPCRs
Species:
Human
Storage Conditions:
Place cells in liquid nitrogen immediately upon receipt. Maintain frozen in liquid nitrogen for up to 5 years. Thaw cells rapidly by removing from liquid nitrogen and immediately immersing in a 37°C water bath. Immediately after ice has thawed, sterilize the exterior of the vial with 70% ethanol. Transfer contents of the vial to a sterile conical tube containing 10 mL growth media, and centrifuge at 200 x g. Remove supernatant and resuspend in fresh growth media. Transfer to a T75 flask containing 20 mL final volume growth media. Cells should be maintained between 0.1 x 106 and 1.0 x 106 cells/mL, and are typically passaged 1:10 every 3-4 days. Cells should be passaged at least once after thawing prior to use in calcium flux assays.