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通过与聚苯乙烯胶乳竞争性结合对低至10纳克/毫升浓度的蛋白质进行定量分析。

Protein quantitation of as low as 10-ng/ml concentrations by competitive binding to polystyrene latexes.

作者信息

Sandwick R K, Schray K J

出版信息

Anal Biochem. 1985 May 15;147(1):210-6. doi: 10.1016/0003-2697(85)90029-6.

Abstract

A protein quantitation method which offers protein detection as low as 10 ng protein/ml and accurate quantitation as low as 30-100 ng protein/ml, depending on the protein, has been designed. The assay, which is relatively quick and simple to perform, utilizes the strong, nonspecific adsorption of proteins onto polystyrene latexes. A competition is created between a marker enzyme and the analyte protein for a limited amount of latex surface area. Due to inactivation of the enzyme upon binding to a hydrophobic latex surface, measurement of enzyme activity allows determination of the bound/free enzyme ratio and thus the competing protein concentration. Considerations of sensitivity and simplicity are suggested to make this assay superior to others presently available.

摘要

已经设计出一种蛋白质定量方法,该方法能够检测低至10 ng蛋白质/毫升的蛋白质,并根据蛋白质种类实现低至30 - 100 ng蛋白质/毫升的准确定量。该检测方法操作相对快速且简单,利用了蛋白质在聚苯乙烯乳胶上的强烈非特异性吸附。标记酶和分析物蛋白质会竞争有限的乳胶表面积。由于酶与疏水性乳胶表面结合后会失活,通过测量酶活性可以确定结合/游离酶的比例,从而确定竞争蛋白的浓度。考虑到灵敏度和简便性,该检测方法被认为优于目前现有的其他方法。

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