Department of Medicine, University of Padova, Padova, Italy.
PLoS One. 2012;7(1):e30911. doi: 10.1371/journal.pone.0030911. Epub 2012 Jan 23.
The performance of two-dimensional electrophoresis in conventional gels in Cartesian coordinates (2-DE) vs. polar coordinates (2-PE) is here evaluated. Although 2-DE is performed in much longer Immobiline gels in the first dimension (17 cm) vs. barely 7-cm in 2-PE, an equivalent resolving power is found. Moreover, due to the possibility of running up to seven Immobiline strips in the radial gel format, the reproducibility of spot position is seen to be higher, this resulting in a 20% higher matching efficiency. As an extra bonus, strings of "isobaric" spots (i.e. polypeptides of identical mass with different pI values) are more resolved in the radial gel format, especially in the 10 to 30 kDa region, where the gel area fans out leaving extra space for spot resolution. In conclusion, this novel gel format in the second dimension of 2D gels is seen as an important improvement of this technique, still one of the most popular in proteome analysis.
在此评估了笛卡尔坐标(2-DE)和极坐标(2-PE)下传统凝胶中的二维电泳(2-DE)的性能。尽管在一维中使用长达 17 厘米的 Immobiline 凝胶进行 2-DE,而在 2-PE 中仅使用 7 厘米,但发现了相当的分辨率。此外,由于在径向凝胶形式中可以运行多达七个 Immobiline 条带,因此斑点位置的重现性更高,这导致匹配效率提高了 20%。作为额外的好处,“等电点”(即具有不同等电点的相同质量的多肽)的“等电点”串在径向凝胶形式中得到了更好的分辨,尤其是在 10 到 30 kDa 区域,其中凝胶区域展开,为斑点分辨率留出了额外的空间。总之,这种 2D 凝胶中二维凝胶的新型凝胶格式被认为是该技术的重要改进,仍然是蛋白质组分析中最受欢迎的技术之一。