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优化 Cydex 蓝法:一种使用环糊精的一步比色法蛋白质测定法,与去污剂和还原剂兼容。

Optimization of the cydex blue assay: A one-step colorimetric protein assay using cyclodextrins and compatible with detergents and reducers.

机构信息

Chemistry and Biology of Metals, UMR5249, Univ. Grenoble Alpes, CNRS, CEA, BIG-LCBM, Grenoble, France.

出版信息

PLoS One. 2018 Apr 11;13(4):e0195755. doi: 10.1371/journal.pone.0195755. eCollection 2018.

Abstract

Sodium dodecyl sulfate electrophoresis (SDS) is a protein separation technique widely used, for example, prior to immunoblotting. Samples are usually prepared in a buffer containing both high concentrations of reducers and high concentrations of SDS. This conjunction renders the samples incompatible with common protein assays. By chelating the SDS, cyclodextrins make the use of simple, dye-based colorimetric assays possible. In this paper, we describe the optimization of the assay, focussing on the cyclodextrin/SDS ratio and the use of commercial assay reagents. The adaptation of the assay to a microplate format and using other detergent-containing conventional extraction buffers is also described.

摘要

十二烷基硫酸钠电泳(SDS)是一种广泛应用的蛋白质分离技术,例如在免疫印迹之前。样品通常在含有高浓度还原剂和高浓度 SDS 的缓冲液中制备。这种组合使样品与常见的蛋白质测定方法不兼容。通过螯合 SDS,环糊精使简单的、基于染料的比色测定成为可能。本文描述了该测定法的优化,重点是环糊精/SDS 比例和使用商业测定试剂。还描述了该测定法向微孔板格式的改编以及使用其他含去污剂的常规提取缓冲液。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a451/5895047/9508f162b914/pone.0195755.g001.jpg

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