A flexible single-step detection of blotted antigen using a fusion protein between protein A and green fluorescent protein.
A green fluorescent protein mutant (S147P GFP) was fused with protein A and expressed in Escherichia coli. This fusion protein (PA-GFP147) was used in immunoblotting studies as a new detection system, designated as "flexible single-step detection (FSSD)". In FSSD, the detection of blotted antigen was done in one step, and the antigen-antibody reaction can be monitored by UV-irradiation in real time. The reaction time, therefore, can be flexibly controlled by monitoring the green fluorescence.
Aoki
T
T
Department of Biochemistry, Faculty of Pharmaceutical Sciences, Health Sciences University of Hokkaido, Ishikari-Tobetsu, Japan. aokit@hoku-iryo-u.ac.jp
Miyashita
M
M
Fujino
H
H
Watabe
H
H
D000818:Animals; D000937:Antigen-Antibody Reactions; D000941:Antigens / Q000276:immunology*; D015153:Blotting, Western / Q000379:methods; D049452:Green Fluorescent Proteins; D006801:Humans; D007124:Immunoenzyme Techniques; D007074:Immunoglobulin G / Q000276:immunology; D008164:Luminescent Proteins / Q000235:genetics / Q000276:immunology*; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology; D013050:Spectrometry, Fluorescence / Q000379:methods; D013205:Staphylococcal Protein A / Q000235:genetics / Q000276:immunology*; D000509:alpha-Fetoproteins / Q000235:genetics / Q000276:immunology*
D000941:Antigens; D007074:Immunoglobulin G; D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; D013205:Staphylococcal Protein A; D000509:alpha-Fetoproteins; D049452:Green Fluorescent Proteins