Senesi S, Freer G, Gasperini M, Angioni M R, Campa M
J Immunol Methods. 1987 Jun 26;100(1-2):197-209. doi: 10.1016/0022-1759(87)90190-6.
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.
本文描述了一种从小鼠淋巴细胞中实现膜结合5'-核苷酸酶（5'-N）良好回收和部分纯化的简单方法。实验步骤基于在聚阳离子珠上分离质膜，并从珠结合的质膜中选择性增溶酶活性。采用这种方法，全细胞匀浆中可检测到的5'-N活性的95%以上可常规地在单一馏分中回收，该馏分显示出5'-N比活性，其比粗匀浆中发现的比活性至少高60倍。该方法还能将5'-N与膜结合碱性磷酸酶（AP）以及任何其他干扰性非特异性磷酸酶完全分离。由于即使在仅有少量淋巴细胞时该方法也快速且高度可重复，因此它可能有助于检测不同T和B淋巴细胞亚群中5'-N活性的变化。
