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  • Title: Mouse lymphocyte enzymatic markers. A rapid method for achieving selective solubilization and efficient recovery of the membrane-bound 5'-nucleotidase.
    Author: Senesi S, Freer G, Gasperini M, Angioni MR, Campa M.
    Journal: J Immunol Methods; 1987 Jun 26; 100(1-2):197-209. PubMed ID: 3036949.
    Abstract:
    A simple method is described for achieving a good recovery and a partial purification of the membrane-bound 5'-nucleotidase (5'-N) from mouse lymphocytes. The experimental procedure is based upon plasma membrane isolation on polycationic beads and selective solubilization of the enzyme activity from bead-bound plasma membranes. With this method, more than 95% of the 5'-N activity detectable in the whole cell homogenates can be routinely recovered in a single fraction showing a 5'-N specific activity which is at least 60 times higher than that found in the crude homogenate. This method also provides a complete separation of 5'-N from the membrane-bound alkaline phosphatase (AP), as well as from any other interfering non-specific phosphatase. Since this method is rapid and highly reproducible even when small amounts of lymphocytes are available, it may be useful for detecting changes in 5'-N activity in the different T- and B-lymphocyte subpopulations.
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