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

Research output: Contribution to journalArticle

21 Citations (Scopus)

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 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).

Original languageEnglish
Pages (from-to)1730-1736
Number of pages7
JournalElectrophoresis
Volume24
Issue number11
DOIs
Publication statusPublished - 2003 Jun 1

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Capillary electrophoresis
Capillary Electrophoresis
Sodium Dodecyl Sulfate
Borates
Tromethamine
Lasers
Polyethylene oxides
Fluorescence
Buffers
Caseins
Proteins
Ethylene Oxide
Injections
Joule heating
Bovine Serum Albumin
Heating
Electrolytes
Milk
Salts

All Science Journal Classification (ASJC) codes

  • Analytical Chemistry
  • Biochemistry
  • Clinical Biochemistry

Cite this

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title = "A simple, rapid, and sensitive method for analysis of SYPRO red labeled sodium dodecyl sulfate-protein complexes by capillary electrophoresis with laser-induced fluorescence",
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 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).",
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A simple, rapid, and sensitive method for analysis of SYPRO red labeled sodium dodecyl sulfate-protein complexes by capillary electrophoresis with laser-induced fluorescence. / Chiu, Tai Chia; Lin, Yang Wei; Huang, Chih Ching; Chrambach, Andreas; Chang, Huan Tsung.

In: Electrophoresis, Vol. 24, No. 11, 01.06.2003, p. 1730-1736.

Research output: Contribution to journalArticle

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T1 - A simple, rapid, and sensitive method for analysis of SYPRO red labeled sodium dodecyl sulfate-protein complexes by capillary electrophoresis with laser-induced fluorescence

AU - Chiu, Tai Chia

AU - Lin, Yang Wei

AU - Huang, Chih Ching

AU - Chrambach, Andreas

AU - Chang, Huan Tsung

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AB - 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).

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