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HTS052M - ChemiScreen™ A3 Adenosine Receptor Membrane Preparation

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HTS052M
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    Extracellular adenosine mediates a multitude of biological effects, including wakefulness, anti-arrythmia, bronchoconstriction and response to ischemia and oxidative stress. A family of four GPCR adenosine receptors, A1, A2A, A2B and A3, is responsible for these effects (Fredholm et al., 2001). A3, which couples to Gi/o, is expressed in mast cells along with A2B. Mice lacking A3 display reduced mast cell degranulation and bronchoconstriction in response to adenosine (Tilley et al., 2003; Zhong et al., 2003). Cloned human A3-expressing cell line is made in the Chem-3 host, which supports high levels of recombinant A3 expression on the cell surface and contains high levels of the promiscuous G protein Ga15 to couple the receptor to the calcium signaling pathway. Thus, the cell line is an ideal tool for screening for agonists and antagonists at A3. The A3 membrane preparations are crude membrane preparations made from our proprietary stable recombinant cell lines to ensure high-level of GPCR surface expression; thus, they are ideal HTS tools for screening of antagonists of A3 interactions with its ligands. The membrane preparations exhibit a Kd of 4.7 nM for [125I]-AB-MECA. With 5mg/well A3 Membrane Prep and 5nM [125I]-AB-MECA, a greater than 3-fold signal-to-background ratio is obtained.

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    Item Unit of Measure: EA
    Quantity: 200 units
    Storage: On receipt of material store at -70°C. Unopened reagent is stable for a minimum of 1 year from date of shipment when stored at recommended storage temperature. Avoid repeat freeze/thaw cycles. For maximum recovery of product, centrifuge original vial
    Applications: Radioligand Binding Assay
    Species: Human A3 Adenosine Receptor (Accession number L22607)
    Host Cell: Chem-3, a suspension mammalian cell line without any endogenous A3 expression.
    Reference 1: 1. Fredholm, BB et al. (2001) International Union of Pharmacology. XXV. Nomenclature and classification of adenosine receptors. Pharmacol. Rev. 53: 527-552.
    Reference 2: 2. Tilley SL et al. (2003) Identification of A3 receptor- and mast cell-dependent and -independent components of adenosine-mediated airway responsiveness in mice. J. Immunol. 2003 Jul 1-171(1):331-7.
    Reference 3: 3. Zhong H et al. (2003) Activation of murine lung mast cells by the adenosine A3 receptor. J. Immunol. 171: 338-45.
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