Stable expression of human A.sub.2B adenosine receptors, and assays employing the same
AssigneeThe University of Patent Foundation
InventorsJoel Linden, Heidi Taylor, Anna Robeva, Robin Woodard, Xiaowei Jin
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Abstract
Stable over-expression of native and hexahistidine/FLAG extended recombinant human adenosine receptors permits recovery of membranes and membrane fragments bearing a high density of adenosine receptors. Cell lines expressing all four sub-types of human adenosine receptors have been identified. The receptors, and membranes and membrane fragments bearing the same, can be used in assays to screen compounds for binding ability to the adenosine receptors, such as potential pharmaceutical agents.
Claims7
A method of detecting binding of .sup.3 H!1,3-diethyl-8-phenyl-xanthine to a human A.sub.2B adenosine receptor, comprising contacting said receptor, which is present in an amount of at least about 5 pmol/mg of protein, with said .sup.3 H!1,3-diethyl-8-phenyl-xanthine under conditions which permit binding of said .sup.3 H!1,3-diethyl-8-phenyl-xanthine to said A.sub.2B adenosine receptor to occur, washing said adenosine receptor to remove any unbound material, and inspecting the resulting sample to determine the presence of said .sup.3 H!1,3-diethyl-8-phenyl-xanthine, wherein the presence and amount of said .sup.3 H!1,3-diethyl8-phenyl-xanthine correlates with the presence and amount of binding of said 3H!1,3 -diethyl-8-phenyl-xanthine to said A.sub.2B adenosine receptor.
A method of detecting binding of .sup.3 H!1,3-diethyl-8-phenyl-xanthine to a human A.sub.2B adenosine receptor, comprising contacting a membrane with said .sup.3 H!1,3-diethyl-8-phenyl-xanthine, wherein said membrane is comprised of a plurality of said A.sub.2B adenosine receptors which are present in an amount of at least about 5 pmol/mg of protein, under conditions which permit binding of said .sup.3 H!1,3-diethyl-8-phenyl-xanthine to said A.sub.2B adenosine receptors to occur, and detecting binding of said .sup.3 H!1,3-diethyl-8-phenyl-xanthine to said A.sub.2B adenosine receptors.
Dependencies are inferred from the claim text.