Alamethicin or detergent permeabilization of the cell membrane as a tool for adenylate cyclase determination: Application to the study of hormone responsiveness in lymphocytes |
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Authors: | Jean-Claude Bonnafous Jacques Dornand Jean-Claude Mani |
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Affiliation: | Laboratoire de Biochimie des Membranes, ER CNRS 228, ENSCM, 8 rue de l''Ecole Normale, F-34075 Montpellier France |
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Abstract: | Treatment of mouse lymphocytes with very low concentrations of alamethicin or Lubrol PX induces spontaneous permeabilization of the plasma membrane to ATP and allows determination of adenylate cyclase activity in whole cells. The permeabilized cells retain responsiveness to hormones (isoproterenol, adenosine analogs) and to fluoride. The main advantage of this new method is that it does not require any homogenization step, and thus adenylate cyclase activities can be accurately and reproducibly measured with very low amounts of cells. It should be especially useful for the study of purified lymphocyte subpopulations. |
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Keywords: | Alamethicin Detergent permeabilization Adenylate cyclase assay (Lymphocyte) |
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