Tags

Type your tag names separated by a space and hit enter

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. 2003 Jun; 24(11):1730-6.E

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 0.45 +/- 0.03 mM (n = 5).

Authors+Show Affiliations

Department of Chemistry, National Taiwan University, Taipei, Taiwan, R.O.C.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

12783449

Citation

Chiu, Tai-Chia, et al. "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, vol. 24, no. 11, 2003, pp. 1730-6.
Chiu TC, Lin YW, Huang CC, et al. 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. 2003;24(11):1730-6.
Chiu, T. C., Lin, Y. W., Huang, C. C., Chrambach, A., & Chang, H. T. (2003). 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, 24(11), 1730-6.
Chiu TC, et al. 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. 2003;24(11):1730-6. PubMed PMID: 12783449.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR 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, PY - 2003/6/5/pubmed PY - 2004/2/11/medline PY - 2003/6/5/entrez SP - 1730 EP - 6 JF - Electrophoresis JO - Electrophoresis VL - 24 IS - 11 N2 - 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 0.45 +/- 0.03 mM (n = 5). SN - 0173-0835 UR - https://www.unboundmedicine.com/medline/citation/12783449/A_simple_rapid_and_sensitive_method_for_analysis_of_SYPRO_Red_labeled_sodium_dodecyl_sulfate_protein_complexes_by_capillary_electrophoresis_with_laser_induced_fluorescence_ DB - PRIME DP - Unbound Medicine ER -