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Drug-induced Sensitization of Adenylyl Cyclase: Assay Streamlining and Miniaturization for Small Molecule and siRNA Screening Applications
Published on: January 27, 2014
Assays of recombinant adenylyl cyclases expressed in Sf9 cells
1Department of Pharmacology, University of Texas Southwestern Medical Center, Dallas, USA.
Abstract:
This chapter outlines procedures for the expression of mammalian membrane-bound adenylyl cyclases (AC) in Sf9 cells and subsequent in vitro methods for assessing the activity of these cyclases. Membrane preparations derived from this overexpression system provide homogeneous sources of mammalian AC because AC that are endogenously expressed in Sf9 cells contribute low amounts of activity relative to the overexpressed enzyme. These approaches are applicable to all nine isoforms of mammalian membrane-bound AC isoforms.

