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使用基于去污剂的缓冲液可以在免疫测定形式中检测到前体抑制素形式。

Use of detergent-based buffers allows detection of precursor inhibin forms in an immunoassay format.

机构信息

Prince Henry's Institute of Medical Research, Clayton, Victoria 3168, Australia.

出版信息

Mol Cell Endocrinol. 2013 Dec 5;381(1-2):106-14. doi: 10.1016/j.mce.2013.07.022. Epub 2013 Jul 30.

Abstract

Inhibin ELISAs are used in monitoring aspects of reproductive function, however these assays are based on the measurements of the mature 30kDa inhibin forms and not precursor forms. In conventional ELISA formats, the 105kDa unprocessed 'Pro-inhibin' forms are immunologically inactive, but the immunoactivity can be recovered in the presence of detergents. The immunoactivity of Pro-inhibin forms was assessed in the presence of a range of detergents utilising antibodies to the α-, βA- and βB-subunits of inhibin. In contrast to mature forms, unprocessed inhibin forms showed a 10-40 fold increase in inhibin A and total inhibin immunoactivities under optimised detergent (0.5% SDS/2% Triton X-100) conditions. The suppressed immunoactivity of the Pro-inhibin forms in these immunoassays was attributed to steric hindrance by the respective βA- and α-subunit prodomains. This study details a detergent-based immunoassay that allows detection of previously undetectable precursor inhibin forms.

摘要

抑制素 ELISA 用于监测生殖功能的各个方面,然而这些检测方法基于对成熟的 30kDa 抑制素形式而不是前体形式的测量。在传统的 ELISA 格式中,未经处理的 105kDa“前抑制素”形式在免疫学上是无活性的,但在去污剂的存在下可以恢复其免疫活性。利用针对抑制素的 α-、βA-和 βB-亚基的抗体,在一系列去污剂存在的情况下评估了前抑制素形式的免疫活性。与成熟形式相比,未经处理的抑制素形式在优化的去污剂(0.5% SDS/2% Triton X-100)条件下,抑制素 A 和总抑制素免疫活性增加了 10-40 倍。前抑制素形式在这些免疫测定中受到抑制的免疫活性归因于各自的 βA-和 α-亚基前导域的空间位阻。本研究详细介绍了一种基于去污剂的免疫测定法,该方法可检测以前无法检测到的前体抑制素形式。

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