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Capillary electrophoresis with laser-induced fluorescence detection of indigo carmine and indigo carmine-labeled proteins.
J Capill Electrophor Microchip Technol. 2002 Sep-Dec; 7(5-6):95-101.JC

Abstract

Indigo carmine blue is a water-soluble, light-sensitive, anionic dye most widely applied in microscopic staining techniques. It is virtually nonfluorescent in its oxidized state in aqueous solution, but when reduced under alkaline conditions, it becomes fluorescent, with absorption and emission maxima at 436 and 528 nm, respectively. It is demonstrated that the fluorescent character of the reduced form of the dye can be exploited as a label for the determination of cationic proteins by capillary electrophoresis with laser-induced fluorescence detection. Model proteins trypsinogen and cytochrome c are noncovalently labeled with indigo carmine; the bound species are reduced, rendering the indigo carmine fluorescent; and capillary electrophoresis with laser-induced fluorescence detection is used for subsequent analysis. Various buffer systems and buffer additives were examined and optimized, and a suitable pH range for optimal fluorescence intensity and protein-dye interaction was established. Fluorescence quenching of the reduced dye when bound to protein was observed in all buffer systems.

Authors+Show Affiliations

Department of Chemistry, Wake Forest University, Winston-Salem, NC 27109, USA.No affiliation info availableNo affiliation info available

Pub Type(s)

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

Language

eng

PubMed ID

12546157

Citation

McCorquodale, Elizabeth Moody, et al. "Capillary Electrophoresis With Laser-induced Fluorescence Detection of Indigo Carmine and Indigo Carmine-labeled Proteins." Journal of Capillary Electrophoresis and Microchip Technology, vol. 7, no. 5-6, 2002, pp. 95-101.
McCorquodale EM, Piper J, Colyer CL. Capillary electrophoresis with laser-induced fluorescence detection of indigo carmine and indigo carmine-labeled proteins. J Capill Electrophor Microchip Technol. 2002;7(5-6):95-101.
McCorquodale, E. M., Piper, J., & Colyer, C. L. (2002). Capillary electrophoresis with laser-induced fluorescence detection of indigo carmine and indigo carmine-labeled proteins. Journal of Capillary Electrophoresis and Microchip Technology, 7(5-6), 95-101.
McCorquodale EM, Piper J, Colyer CL. Capillary Electrophoresis With Laser-induced Fluorescence Detection of Indigo Carmine and Indigo Carmine-labeled Proteins. J Capill Electrophor Microchip Technol. 2002 Sep-Dec;7(5-6):95-101. PubMed PMID: 12546157.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Capillary electrophoresis with laser-induced fluorescence detection of indigo carmine and indigo carmine-labeled proteins. AU - McCorquodale,Elizabeth Moody, AU - Piper,Jonathan, AU - Colyer,Christa L, PY - 2003/1/28/pubmed PY - 2003/7/30/medline PY - 2003/1/28/entrez SP - 95 EP - 101 JF - Journal of capillary electrophoresis and microchip technology JO - J Capill Electrophor Microchip Technol VL - 7 IS - 5-6 N2 - Indigo carmine blue is a water-soluble, light-sensitive, anionic dye most widely applied in microscopic staining techniques. It is virtually nonfluorescent in its oxidized state in aqueous solution, but when reduced under alkaline conditions, it becomes fluorescent, with absorption and emission maxima at 436 and 528 nm, respectively. It is demonstrated that the fluorescent character of the reduced form of the dye can be exploited as a label for the determination of cationic proteins by capillary electrophoresis with laser-induced fluorescence detection. Model proteins trypsinogen and cytochrome c are noncovalently labeled with indigo carmine; the bound species are reduced, rendering the indigo carmine fluorescent; and capillary electrophoresis with laser-induced fluorescence detection is used for subsequent analysis. Various buffer systems and buffer additives were examined and optimized, and a suitable pH range for optimal fluorescence intensity and protein-dye interaction was established. Fluorescence quenching of the reduced dye when bound to protein was observed in all buffer systems. SN - 1946-4940 UR - https://www.unboundmedicine.com/medline/citation/12546157/Capillary_electrophoresis_with_laser_induced_fluorescence_detection_of_indigo_carmine_and_indigo_carmine_labeled_proteins_ DB - PRIME DP - Unbound Medicine ER -