A simple, rapid, and sensitive method for analysis of SYPRO red labeled sodium dodecyl sulfate-protein complexes by capillary electrophoresis with laser-induced fluorescence

Tai Chia Chiu, Yang Wei Lin, Chih Ching Huang, Andreas Chrambach, Huan Tsung Chang

研究成果: Article

21 引文 斯高帕斯(Scopus)

摘要

We describe a segmental filling method for the analysis of SYPRO Red labeled sodium dodecyl sulfate (SDS)-proteins (SRSPs) by capillary electrophoresis-laser induced fluorescence (CE-LIF) with electroosmotic counterflow of poly(ethylene oxide) (PEO). It is shown that SDS and salt play a crucial role in determining the fluorescence intensity of the SRSP Although the fluorimetric measurements reveal that the SRSPs fluoresce strongly in Tris-borate (TB) buffer containing 0.1 % SDS and high concentrations of NaCl (100 mM), these conditions are not appropriate to CE in view of Joule heating. To overcome that impediment, we applied a plug of 0.1 % SDS (1/5 to 1/3 of the injection volume) prior to injection of samples (0.64 μL) prepared in TB buffer containing 50 mM NaCl and SYPRO Red. When using a background electrolyte of 0.6% PEO in TB buffer containing NaCl, electroosmotic counterflow of the analytes allows one to concentrate large sample volumes (up to 1/3 of effective capillary length) in 21 min, with detection of 0.35 and 0.10 nM for bovine serum albumin and casein, respectively. With a linear dynamic range from 10 nM to 5 μM this method provides the capability of determining the concentration of casein in cow's milk as 0.45 ± 0.03 mM (n = 5).

原文English
頁(從 - 到)1730-1736
頁數7
期刊Electrophoresis
24
發行號11
DOIs
出版狀態Published - 2003 六月 1

    指紋

All Science Journal Classification (ASJC) codes

  • Analytical Chemistry
  • Biochemistry
  • Clinical Biochemistry

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