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

Electrophoresis
Tai-Chia ChiuHuan-Tsung Chang

Abstract

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 microL) 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 microM, this method provides the capability of determining the concentration of casein in cow's milk as...Continue Reading

Citations

Nov 10, 2007·Journal of Chromatography. a·Yi ChenChanggui Qiu
Dec 1, 2006·Electrophoresis·Ana M García-CampañaHuguette Fabre
Nov 25, 2005·Electrophoresis·Vladislav Dolník
Aug 24, 2006·Electrophoresis·Yu-Fen HuangHuan-Tsung Chang
Nov 30, 2011·Analytica Chimica Acta·Zaifang ZhuShaorong Liu
May 24, 2006·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Maria Rowena N Monton, Shigeru Terabe
Feb 8, 2006·Analytical Biochemistry·Alexander V StoyanovSulma Mohammed

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