Pushparajan Charlotte, Goswami Shailesh K, McAdam C John, Hanton Lyall R, Dearden Peter K, Moratti Stephen C, Cridge Andrew G
Department of Biochemistry, University of Otago, Dunedin, New Zealand.
Department of Chemistry, University of Otago, Dunedin, New Zealand.
Electrophoresis. 2018 Mar;39(5-6):824-832. doi: 10.1002/elps.201700303. Epub 2017 Dec 11.
Polyacrylamide gel electrophoresis is a universal tool in a biochemist's toolkit for protein and nucleic acid separation and subsequent visualisation and analysis. The standard formulation of polyacrylamide gels consists of acrylamide (ACM) monomer crosslinked with bisacrylamide (MBA) which creates a gel with excellent sieving properties but which is mechanically fragile and prone to tearing during post-electrophoresis manipulations involved in visualisation and analysis. By adding a poly(ethylene oxide) macro-crosslinker to the standard gel formulation, we have created a tough gel matrix that can be used to fractionate proteins and nucleic acids by polyacrylamide gel electrophoresis. The protein and nucleic acid resolving capabilities and performance during staining and electroblotting of the tough gel matrix rivals that of conventional acrylamide/bisacrylamide gels. The tough gel matrix is resistant to tear and remarkably elastic, capable of stretching to over four times its original length before breaking, and represents a significant improvement over standard polyacrylamide gel formulations.
聚丙烯酰胺凝胶电泳是生物化学家工具包中用于蛋白质和核酸分离以及后续可视化和分析的通用工具。聚丙烯酰胺凝胶的标准配方由丙烯酰胺(ACM)单体与双丙烯酰胺(MBA)交联而成,这种凝胶具有出色的筛分特性,但机械性能脆弱,在可视化和分析过程中涉及的电泳后操作中容易撕裂。通过向标准凝胶配方中添加聚环氧乙烷大交联剂,我们创建了一种坚韧的凝胶基质,可用于通过聚丙烯酰胺凝胶电泳分离蛋白质和核酸。这种坚韧凝胶基质在染色和电转印过程中的蛋白质和核酸分辨能力及性能可与传统丙烯酰胺/双丙烯酰胺凝胶相媲美。该坚韧凝胶基质抗撕裂且具有显著的弹性,能够在断裂前拉伸至其原始长度的四倍以上,代表了对标准聚丙烯酰胺凝胶配方的重大改进。