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首页> 外文期刊>Electrophoresis: The Official Journal of the International Electrophoresis Society >High-resolution separation of peptides by sodium dodecyl sulfate-polyacrylamide gel 'focusing'.
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High-resolution separation of peptides by sodium dodecyl sulfate-polyacrylamide gel 'focusing'.

机译:通过十二烷基硫酸钠-聚丙烯酰胺凝胶“聚焦”进行肽的高分辨率分离。

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摘要

As a follow-up of our previous report (Anal. Chem. 2007, 79, 821-827) on analytical SDS-PAGE focusing, a refinement of the method for separation of peptides in the small to medium M(r) range (0.5-10 kDa) is here reported, based on a shallow gradient of immobilized positive charges (0-10 mM) onto a minimally sieving polyacrylamide gel matrix (4%T, 2.5%C). Unlike conventional SDS-PAGE, which rarely can achieve the separations of polypeptide chains below a critical value of 10 kDa, the present method can be fine-tuned to perform such separations even down to a size of only 500 Da. In the case of larger fragments, the major peptide zones are shown, under microscope observation, to be composed by envelops of bands as narrow as 20-100 mum, spaced at regular intervals of 100-150 mum. It is hypothesized that such larger peptides could form complexes with rather small SDS micelles and that such peptide-SDS complexes could differ in charge by just a single negative charge.
机译:作为我们先前关于分析SDS-PAGE聚焦的报告(Anal。Chem。2007,79,821-827)的后续报告,对中小M(r)范围(0.5)肽段的分离方法进行了改进根据固定的正电荷(0-10 mM)在最小筛分的聚丙烯酰胺凝胶基质(4%T,2.5%C)上的浅梯度,本文报道了-10 kDa)。与常规的SDS-PAGE不同,常规的SDS-PAGE很少能实现低于10 kDa的临界值的多肽链的分离,本发明的方法可以进行微调以进行这样的分离,甚至可以减小到只有500 Da的大小。在较大的片段的情况下,在显微镜观察下,主要的肽区域显示为由窄至20-100微米的条带包围,以100-150微米的规则间隔隔开。假设这样的较大的肽可以与相当小的SDS胶束形成复合物,并且这样的肽-SDS络合物的电荷可以仅有一个负电荷而不同。

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