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抗人血栓素合酶多克隆抗体和单克隆抗体的制备与鉴定

Production and characterization of polyclonal and monoclonal antibodies against human thromboxane synthase.

作者信息

Nüsing R, Wernet M P, Ullrich V

机构信息

Faculty of Biology, University of Konstanz, FRG.

出版信息

Blood. 1990 Jul 1;76(1):80-5.

PMID:2364175
Abstract

Polyclonal and monoclonal antibodies (MoAbs) were raised against human platelet thromboxane (Tx) synthase. Neither the antiserum nor the MoAbs inhibited the enzyme activity significantly. Three MoAbs, Tü 300, Kon 6, and Kon 7, were purified and further characterized. They are monospecific as shown by activity precipitation or Western blot analysis, and recognized different epitopes on Tx-synthase. Tü 300 could precipitate the enzyme and recognized conformational epitopes, whereas Kon 6 and Kon 7 only reacted in Western blots. Antibody Tü 300 can be used in immunohistology but shows no crossreactivity with Tx-synthase from other species. In human lung tissue staining with peroxidase, coupled Tü 300 was only found in alveolar macrophages.

摘要

制备了针对人血小板血栓素(Tx)合酶的多克隆抗体和单克隆抗体(MoAb)。抗血清和单克隆抗体均未显著抑制该酶的活性。对三种单克隆抗体Tü 300、Kon 6和Kon 7进行了纯化和进一步鉴定。通过活性沉淀或蛋白质免疫印迹分析表明它们具有单特异性,并且识别Tx合酶上不同的表位。Tü 300能沉淀该酶并识别构象表位,而Kon 6和Kon 7仅在蛋白质免疫印迹中发生反应。抗体Tü 300可用于免疫组织学研究,但与其他物种的Tx合酶无交叉反应性。在用过氧化物酶进行染色的人肺组织中,偶联的Tü 300仅在肺泡巨噬细胞中发现。

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