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Title: A Simple, Rapid, and Sensitive Method for Analysis of SYPRO Red Labeled SDS-Protein Complexes by Capillary Electrophoresis with Laser-Induced Fluorescence
Authors: Chiu, T.-C.;Lin, Y.-W.;Huang, C.-C.;Chrambach, A.;Chang, H.-T.
Contributors: 化學系
Keywords: Capillary electrophoresis
Laser-induced fluorescence
Poly(ethylene oxide)
Sodium dodecyl sulfate-proteins
Date: 2003
Issue Date: 2010-11-10T06:26:30Z
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 L) 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 M, this method provides the capability of determining the concentration of casein in cow’s milk as 0.45 0.03 mM (n = 5).
Relation: Electrophoresis, 24, 1730-1736
Appears in Collections:[Department of Chemistry] Periodical Articles

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