Stable, recombinant HEK293 cell line expressing human TMEM16A (transmembrane protein 16A, also called anoctamin 1, calcium-activated chloride channel (ANO1), accession number NM_018043).
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Synonyms
TMEM16A, transmembrane protein 16A, anoctamin 1, calcium-activated chloride channel, ANO1
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Product Data Gallery
TMEM16A expressed in HEK293 produced I- influx after extracellular addition of I- with ionomycin. A) TMEM16A-HEK293 cells; B) WT-HEK293 cells
Ionomycin-induced I- influx in TMEM16A-HEK293 cells was blocked by niflumic acid, a CaCC channel blocker
Each vial contains ~1 X 106 cells in 1 ml of 10% DMSO.
Biological Activity
N-terminal FLAG tagged human TMEM16A channel has been stably expressed in HEK293 cell line and its expression was confirmed by Western blotting. The CaCC activity of TMEM16A was characterized by an assay based on halide-sensitive yellow fluorescent protein (YFP) mutant whose fluorescence is quenched by increasing halide concentration. When TMEM16A-expressed HEK293 cells were stimulated with ionomycin to raise the intracellular level of Ca2+, TMEM16A produced I– influx in HEK293 that triggered the rapid decrease
of fluorescence of transfected YFP mutant. The ionomycin-induced I– influx through TMEM16A was blocked by niflumic acid, a CaCC channel blocker.
Genbank #
NM_018043
UniProt #
Q5XXA6
Mycoplasma Testing
The cell line has been screened using the PCR-based VenorGeM Mycoplasma Detection kit (Sigma Aldrich) to confirm the absence of Mycoplasma species.
Background
Calcium-activated chloride channels (CaCCs) are major regulators in numerous physiological processes including sensory transduction, epithelial secrection, cardiac and neutonal excitation, and smooth muscle contraction.
TMEM16A (ANO1), a member of a family of putative plasma membrane proteins, is identified as a CaCC that is activated by intracellular Ca2+ and Ca2+-mobilizing stimuli. It has eight putative transmembrane segments without domains evidently involved in calcium regulation. The relative permeability of TMEM16A to monovalent anions is NO3- > I- > Br- > Cl- > F-.
References
1. Verkman A.S. and Galietta L.J.V. (2009) Cholide channels as drug targets. Nature Reviews8: 153-171. 2. Caputo A. et al. (2008). Science322: 590-594. 3. Yang Y.D. et al. (2008). Nature455: 1210-1215.
Storage/Stability
Store in liquid nitrogen immediately upon receipt.
Growth Media
For best results, it is highly recommended to use these validated and optimized media from BPS Bioscience. Other preparations or formulations of media may result in suboptimal performance.
Thaw Medium 1 (BPS Cat. #60187): MEM medium supplemented with 10% FBS, 1% non-essential amino acids, 1 mM Na pyruvate, 1% Penicillin/Streptomycin
Growth Medium 1B (BPS Cat. #79531): MEM medium supplemented with 10% FBS, 1% non-essential amino acids, 1 mM Na pyruvate, 1% Penicillin/Streptomycin plus 400 μg/ml of Geneticin to ensure the recombinant expression is maintained.
Instructions for Use
See data sheet for detailed culturing and assay protocol.
Applications
• Drug compound screening for activators or inhibitors of TMEM16A • Functional assays for TMEM16A • Efficient antigen for mouse immunization
Shipping Temperature
-80°C
Notes
License Disclosure Visit bpsbioscience.com/license for the label license and other key information about this product.
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