ChemiScreen™ alpha2A Calcium-Optimized FLIPR Cell Lines
Description:
ChemiScreen™ alpha2A Calcium-Optimized FLIPR Cell Lines
Trade Name:
ChemiScreen
Product Overview:
TRANSFECTION: Full-length human ADRA2A transcript variant 1 cDNA encoding alpha2A
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 alpha- and beta-adrenoceptors (Bylund et al., 1994). The alpha2 adrenergic receptor subfamily members, consisting of alpha2A, alpha2B, and alpha2C, couple primarily to Gi to inhibit cAMP production, and play an important role in regulation of cardiovascular and CNS function. The alpha2A receptor at presynaptic sites has an inhibitory effect on catecholamine release from sympathetic nerve endings. Experiments with alpha2A-selective agonists and mice lacking alpha2A demonstrate that activation of alpha2A results in hypotension, sedation, analgesia, and hypothermia (Kable et al., 2000). Chemicon's cloned human--2A-expressing cell line is made in the Chem-1 host, which supports high levels of recombinant alpha2A expression on the cell surface and contains high levels of the promiscuous G protein Galpha15 to couple the receptor to the calcium signaling pathway. Thus, the cell line is an ideal tool for screening for antagonists of interactions between-alpha2A and its ligands.
Application Notes:
Calcium flux assay, ligand binding assays
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-1, an adherent cell line expressing the promiscuous G protein, Gα15.
Packaging:
1 mL per vial
Materials Required but Not Delivered:
GROWTH MEDIA: DMEM containing 4.5 g/L glucose/10% heat inactivated fetal bovine serum/1x nonessential amino acids/10 mM HEPES/0.25 mg/ml Geneticin (G418)/100 U/ml each penicillin and streptomycin
Cell Type:
Chem-1
Gene Symbol:
- ADRA2A
- ZNF32
- ADRA2R
- ALPHA2AAR
- Alpha-2AAR
- ADRA2
- OTTHUMP00000058849
- ADRAR
Format:
Cell Line
Presentation:
Cells are frozen at 2 x 106 cells/mL in DMEM/20% fetal bovine serum/100 U/ml penicillin and streptomycin/10% DMSO. Cell line tests negative for mycoplasma.
Quality Assurance:
SPECIFICATIONS: EC50 for calcium mobilization by UK14,304: ~ 2.3 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 37ºC water bath. Immediately after ice has thawed, sterilize the exterior of the vial with 70% ethanol. Transfer contents of the vial to a T75 flask containing 20 mL growth media, and place in a humidified 37°C incubator with 5% CO2.. After 8-24 h, cells will adhere to the plate, at which time the media should be replaced to remove residual DMSO. Cells are passaged by washing with Ca++ and Mg++-free HBSS (10 mL/T75), incubating with 0.05% trypsin/0.2 g/L EDTA (1 mL/T75) for 5-10 minutes at 37°C, and rapping the side of the flask to dislodge the cells. Neutralize the trypsin by addition of 4 volumes growth media. Cells are typically passaged 1:10 with every 3-4 days, and should be passaged at least once after thawing prior to use in calcium flux assays.