一种简单的电动装置，用于从蛋白质消化物中去除十二烷基硫酸钠。
A simple apparatus for electrokinetic removal of sodium dodecyl sulfate from protein digests.
机构信息
Australian Centre for Research on Separation Science (ACROSS), School of Natural Sciences-Chemistry, University of Tasmania, Hobart 7001, Australia.
Central Science Laboratory, Research Division, University of Tasmania, Hobart 7001, Australia.
出版信息
J Chromatogr A. 2020 Sep 27;1628:461443. doi: 10.1016/j.chroma.2020.461443. Epub 2020 Aug 1.
Sodium dodecyl sulfate (SDS) in proteomics samples needs to be removed and estimated prior to mass spectrometry (MS)-based analysis and to avoid MS ion-source contamination. Here, we describe an organic solvent free method to remove SDS using a simple apparatus that mainly consists of an agarose gel inside a 1 mL plastic micropipette tip and a voltage power supply with electrodes. A small volume of sample (e.g., 50 μL) is loaded on top of the gel and then voltage (cathode at the sample side) is applied with an acidic solution at the other end of the micropipette tip. Within 25 min, SDS was removed (e.g., ≥99% SDS in 3.5 mM SDS) and the peptides were retained in the sample solution. The strategy was compared to the commercially available and expensive Pierce spin column for the removal of SDS and recovery of peptides from a digested bovine serum albumin sample.
在基于质谱（MS）的分析之前，需要去除和估计蛋白质组学样品中的十二烷基硫酸钠（SDS），以避免 MS 离子源的污染。在这里，我们描述了一种无有机溶剂的方法，使用一种简单的装置来去除 SDS，该装置主要由一个琼脂糖凝胶和一个带有电极的 1 mL 塑料微量移液器组成。将少量样品（例如 50 μL）加载在凝胶顶部，然后在微量移液器的另一端施加酸性溶液以施加电压（阴极在样品侧）。在 25 分钟内，SDS 被去除（例如，在 3.5 mM SDS 中≥99% SDS），并且肽保留在样品溶液中。该策略与商业上可获得的昂贵的 Pierce 螺旋柱进行了比较，用于从消化的牛血清白蛋白样品中去除 SDS 和回收肽。
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