Highly sensitive double-fluorescent dye staining on microchip electrophoresis for analysis of milk proteins

Hiroki Okada, Noritada Kaji, Manabu Tokeshi, Yoshinobu Baba

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

We demonstrated a highly sensitive double-fluorescent dye staining in microchip electrophoresis (ME) for analysis of milk proteins. The detection sensitivity of ME was very limited so far and needed improvement. Our staining method consisted of two steps. First, in sample preparation before electrophoresis, protein was covalently bound to an amine-reactive fluorescent dye, Cy5. Then, the Cy5-attached protein was denatured with SDS and was further stained during electrophoresis, with Agilent fluorescent dye, which was non-covalently attached to hydrophobic regions of the SDS-protein complexes. This double-fluorescent staining enhanced fluorescent intensity and lowered the detection limit to 200 pg of protein. This provided higher sensitivity than silver- or SYPRO Ruby-staining methods, which have previously given the highest sensitivity in protein staining. In addition, we applied our staining method to analysis of milk proteins and achieved their successful detection, whereas it was difficult to analyze them by the unimproved method.

Original languageEnglish
Pages (from-to)2533-2538
Number of pages6
JournalELECTROPHORESIS
Volume29
Issue number12
DOIs
Publication statusPublished - Jun 1 2008

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Microchip Analytical Procedures
Microchip Electrophoresis
Milk Proteins
Electrophoresis
Fluorescent Dyes
Staining and Labeling
Proteins
Ruby
Silver
Amines
Limit of Detection

All Science Journal Classification (ASJC) codes

  • Clinical Biochemistry

Cite this

Highly sensitive double-fluorescent dye staining on microchip electrophoresis for analysis of milk proteins. / Okada, Hiroki; Kaji, Noritada; Tokeshi, Manabu; Baba, Yoshinobu.

In: ELECTROPHORESIS, Vol. 29, No. 12, 01.06.2008, p. 2533-2538.

Research output: Contribution to journalArticle

Okada, Hiroki ; Kaji, Noritada ; Tokeshi, Manabu ; Baba, Yoshinobu. / Highly sensitive double-fluorescent dye staining on microchip electrophoresis for analysis of milk proteins. In: ELECTROPHORESIS. 2008 ; Vol. 29, No. 12. pp. 2533-2538.
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