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Applications: Drug compound screening for activators or inhibitors of TMEM16B
Functional assays for TMEM16B
Efficient antigen for mouse immunization
Background: Calcium-activated chloride channels (CaCCs) are involved in a variety of physiological functions including smooth muscle contraction and olfaction. TMEM16B (ANO2) has been identified as a CaCC that is activated by intracellular Ca2+ and Ca2+-mobilizing stimuli. It has eight putative transmembrane segments and is permeable to monovalent anions.
Biological Activity: Human TMEM16B channel has been stably expressed in HEK293 cell line and its expression was confirmed by Western blotting. The CaCC activity of TMEM16B was characterized by an assay based on a halidesensitive yellow fluorescent protein (YFP) mutant whose fluorescence is quenched by increasing halide concentration. When TMEM16B-expressed HEK293 cells were stimulated with ionomycin to raise the intracellular level of Ca2+, TMEM16B produced I- influx in HEK293, triggering a rapid decrease of fluorescence by the transfected YFP mutant. The ionomycin-induced I- influx through TMEM16B was blocked by niflumic acid, a CaCC channel blocker.
Description: Recombinant HEK293 cell line expressing human TMEM16B (transmembrane protein16B, also called as anoctamin 2, calcium-activated chloride channel (ANO2), accessionnumber NM_001278596).
Genbank: NM_001278596
Host Cell Line: HEK293
Mycoplasma Testing: The cell line has been screened using the PCR-based Venor™GeM MycoplasmaDetection kit (Sigma-Aldrich) to confirm the absence of Mycoplasma species.
Storage Stability: Store in liquid nitrogen immediately upon receipt.
Supplied As: Each vial contains ~1.5 X 10^6 cells in 1 ml of 10% DMSO.
Uniprot: 19475416
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: BSL-2
References: 1. Verkman A.S. and Galietta L.J.V. (2009) Cholide channels as drug targets. NatureReviews 8:153-171.2. Scudieri P. et al. (2012) The anoctamin family: TMEM16A and TMEM16B as calciumactivatedchloride channels. Exp. Physiol. 97(2):177-183.3. Stöhr H. et al. (2009) TMEM16B, a novel protein with calcium-dependent chloridechannel activity, associates with a presynaptic protein complex in photoreceptorterminals. J. Neurosci. 29(21):6809-6818.
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