IMMUNOAFFINITY PURIFICATION OF DNA POLYMERASE-DELTA
IMMUNOAFFINITY PURIFICATION OF DNA POLYMERASE-DELTA
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DOI：
10.1016/0003-9861(95)90013-6
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发表时间：
1995-07-10
影响因子：
3.9
通讯作者：
LEE, MYWT
中科院分区：
文献类型：
--
作者：
JIANG, YQ;ZHANG, SJ;LEE, MYWT
本文报道了一种抗人DNA聚合酶δ（pol δ）的单克隆抗体，其性质适合于免疫亲和层析。高碘酸盐氧化后将抗体固定化并偶联至酰肼活化的支持物。从部分纯化的制备物开始，小牛胸腺pol δ在一个步骤中纯化约200倍。在ssDNA-纤维素上进一步纯化导致分离均匀的制备物。0.3从0.75kg小牛胸腺中分离纯化pol δ的方法比常规方法高约18倍。该方法在分离pol δ方面提供了显著的进步，使其易于从组织中以良好的产率分离。该酶的表征表明，这两个亚基仍然关联的甘油梯度ultracentragation，即使在2.8 M尿素的存在下。(C)出版社：Academic Press
A monoclonal antibody against human DNA polymerase delta (pol delta) was isolated with properties suitable for its utilization for immunoaffinity chromatography. The antibody was immobilized after periodate oxidation and coupled to a hydrazide-activated support. Starting from a partially purified preparation, calf thymus pol delta was purified about 200-fold in a single step, Further purification on ssDNA-cellulose resulted in isolation of a homogeneous preparation, The amount of enzyme isolated, ca. 0.3 mg of pure pol delta from 0.75 kg of calf thymus, is about 18-fold greater than can be achieved by conventional procedures, This procedure provides a significant advance in the isolation of pol delta in allowing its facile isolation from tissues in good yield, The isolated enzyme consisted of two subunits of 125 and 50 kDa. Characterization of the enzyme showed that these two subunits remained associated on glycerol gradient ultracentrifugation even in the presence of 2.8 M urea. (C) 1995 Academic Press, Inc.