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2',5'-寡腺苷酸特异性抗血清的制备、表征及应用

Preparation, characterization, and use of an antiserum specific for 2',5'-oligoadenylates.

作者信息

Hersh C L, Reid T R, Friedman R, Stark G R

出版信息

J Biol Chem. 1984 Feb 10;259(3):1727-30.

PMID:6693433
Abstract

Periodate-oxidized ppp(A2'p)3A was coupled in high yield to an acyl hydrazide derivative of bovine serum albumin and the conjugate was used to immunize rabbits. Several potent and specific antisera were obtained, and one was tested extensively. Fifty per cent of the bound probe ppp(A2'p)3A3'[32P]pCp was displaced by compounds containing the moiety -pA2'pA-at concentrations of 20-60 nM and by compounds containing the moiety -p(A2'p)2A- at 1 nM. The 3',5'-oligoadenylate (A3'p)3A was bound more than 10,000 times less tightly than (A2'p)3A. The antiserum can be used in competition assays in which the amount of an antibody-probe complex is measured after binding to Millipore filters or to polystyrene beads. These assays allow the clear detection of as little as 20 fmol (20 microliters of 1 nM) of 2',5'-oligoadenylates.

摘要

高碘酸盐氧化的ppp(A2'p)3A以高产率与牛血清白蛋白的酰肼衍生物偶联,所得缀合物用于免疫兔子。获得了几种高效且特异的抗血清,并对其中一种进行了广泛测试。在浓度为20 - 60 nM时,含有-pA2'pA-部分的化合物以及在1 nM时含有-p(A2'p)2A-部分的化合物能置换50%结合的探针ppp(A2'p)3A3'[32P]pCp。3',5'-寡腺苷酸(A3'p)3A的结合紧密程度比(A2'p)3A低10000倍以上。该抗血清可用于竞争分析,即先将抗体-探针复合物与微孔滤膜或聚苯乙烯珠结合,然后测量其含量。这些分析能够清晰检测低至20 fmol(20微升1 nM)的2',5'-寡腺苷酸。

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