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绵羊滋养层蛋白-1(来自绵羊孕体的抗黄体溶解蛋白)放射免疫分析方法的建立。

Development of a radioimmunoassay for ovine trophoblast protein-1, the antiluteolytic protein from the sheep conceptus.

作者信息

Vallet J L, Bazer F W, Ashworth C J, Johnson H M, Pontzer C H

机构信息

Department of Animal Science, University of Florida, Gainesville 32611.

出版信息

J Endocrinol. 1988 May;117(2):R5-8. doi: 10.1677/joe.0.117r005.

Abstract

A radioimmunoassay has been developed for quantitation of ovine trophoblast protein-1 (oTP-1), a sheep conceptus secretory protein which allows for maintenance of the corpus luteum during early pregnancy. The assay was validated for dialysed and undialysed culture medium and pregnant uterine flushings ranging from no dilution (neat) to dilutions of 1:2500 for dialysed media, 1:100-1:1000 for undialysed media and 1:50-1:1000 for pregnant uterine flushings. The assay accurately measured oTP-1 added to undiluted and diluted dialysed and undialysed culture media and pregnant uterine flushings. No cross-reaction was detectable for bovine alpha or gamma interferon, bovine calmodulin, feline conceptus secretory proteins, equine conceptus secretory proteins, porcine conceptus secretory proteins, bovine conceptus secretory proteins and proteins in a uterine flushing collected from a non-pregnant ewe. Immunoreactivity in the assay matched that for oTP-1 throughout oTP-1 purification. This assay is the first validated assay which may be used to quantitate production of oTP-1 in culture or content of oTP-1 in uterine flushings.

摘要

已开发出一种放射免疫测定法,用于定量检测绵羊滋养层蛋白-1(oTP-1),这是一种绵羊孕体分泌蛋白,可在妊娠早期维持黄体功能。该测定法已针对透析和未透析的培养基以及妊娠子宫冲洗液进行了验证,透析培养基的稀释范围为不稀释(原液)至1:2500,未透析培养基为1:100 - 1:1000,妊娠子宫冲洗液为1:50 - 1:1000。该测定法能准确测量添加到未稀释和稀释的透析及未透析培养基以及妊娠子宫冲洗液中的oTP-1。对于牛α或γ干扰素、牛钙调蛋白、猫孕体分泌蛋白、马孕体分泌蛋白、猪孕体分泌蛋白、牛孕体分泌蛋白以及从未孕母羊收集的子宫冲洗液中的蛋白质,均未检测到交叉反应。在整个oTP-1纯化过程中,该测定法的免疫反应性与oTP-1的免疫反应性相匹配。此测定法是首个经过验证的测定法,可用于定量检测培养物中oTP-1的产生量或子宫冲洗液中oTP-1的含量。

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