Premium
Study of protein–protein binding reaction by whole‐column fluorescence‐imaged CIEF
Author(s) -
Wu XingZheng,
Asai Shinya,
Yamaguchi Yoshie
Publication year - 2009
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.200800506
Subject(s) - fluorescence , chemistry , bovine serum albumin , protein g , chromatography , binding protein , biophysics , biochemistry , biology , antibody , physics , quantum mechanics , gene , immunology
Whole‐column fluorescence‐imaged CIEF was applied to study protein–protein binding reaction. A homemade whole‐column fluorescence‐imaged CIEF experimental setup was built, and its CIEF performance was evaluated with native fluorescent protein green fluorescence protein and fluorescently labeled proteins (bovine albumin, human albumin, and BSA). p I s, focusing time, detection limits, and linear quantitative range of the proteins were obtained. Furthermore, the method was used to study FITC–protein A–human IgG binding reaction. Experimental results showed that the apparent binding ratio of the FITC–protein A to human IgG was 1:2, and p I of the binding conjugates were about 6.3–6.5. No binding reaction was found between green fluorescence protein and the fluorescent‐labeled proteins.