带碰撞胶电泳。

Band-collision gel electrophoresis.

机构信息

Department of Chemistry and Biochemistry, University of California, Los Angeles, Los Angeles, CA, 90095, USA.

Department of Physics and Astronomy, University of California, Los Angeles, Los Angeles, CA, 90095, USA.

出版信息

Nat Commun. 2019 Aug 12;10(1):3631. doi: 10.1038/s41467-019-11438-9.

DOI:10.1038/s41467-019-11438-9

PMID:31406115

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC6690962/

Abstract

Electrophoretic mobility shift assays are widely used in gel electrophoresis to study binding interactions between different molecular species loaded into the same well. However, shift assays can access only a subset of reaction possibilities that could be otherwise seen if separate bands of reagent species might instead be collisionally reacted. Here, we adapt gel electrophoresis by fabricating two or more wells in the same lane, loading these wells with different reagent species, and applying an electric field, thereby producing collisional reactions between propagating pulse-like bands of these species, which we image optically. For certain pairs of anionic and cationic dyes, propagating bands pass through each other unperturbed; yet, for other pairs, we observe complexing and precipitation reactions, indicating strong attractive interactions. We generalize this band-collision gel electrophoresis (BCGE) approach to other reaction types, including acid-base, ligand exchange, and redox, as well as to colloidal species in passivated large-pore gels.

摘要

电泳迁移率变动分析广泛应用于凝胶电泳中，以研究装入同一泳道的不同分子物种之间的结合相互作用。然而，迁移率变动分析只能获取反应的一个子集，如果不同的试剂物种可能发生碰撞反应，则可以看到其他反应可能性。在这里，我们通过在同一泳道中制造两个或更多的小槽，将不同的试剂物种装入这些小槽，并施加电场，从而在这些物种的脉冲状传播带之间产生碰撞反应，我们通过光学成像来观察这些反应。对于某些阴离子和阳离子染料对，传播带可以不受干扰地相互穿过；然而，对于其他染料对，我们观察到络合和沉淀反应，表明存在强烈的吸引力相互作用。我们将这种带碰撞凝胶电泳（BCGE）方法推广到其他反应类型，包括酸碱、配体交换和氧化还原反应，以及钝化大孔凝胶中的胶体物种。