Monitoring of a conjugation reaction between fluorescein isothiocyanate and myoglobin by capillary zone electrophoresis.
The conjugation reaction between the amine reactive fluorescent probe fluorescein isothiocyanate and the protein horse heart myoglobin was directly monitored as the reaction progressed by capillary zone electrophoresis with UV absorbance detection. Kinetic analysis of the data indicated that the order of the reaction was first with respect to myoglobin and 1.3 with respect to fluorescein isothiocyanate. The separation of peaks attributable to the incorporation of n = 1,2, ... 7 fluorochrome labels into the protein has some significance for the generation of single-label protein probes, such as antibodies.
Banks
P R
PR
Department of Chemistry and Biochemistry, Concordia University, Montreal, Quebec, Canada.
Paquette
D M
DM
D000818:Animals; D004586:Electrophoresis / Q000379:methods*; D016650:Fluorescein-5-isothiocyanate / Q000737:chemistry*; D006736:Horses; D007700:Kinetics; D009206:Myocardium / Q000737:chemistry; D009211:Myoglobin / Q000737:chemistry*; D013056:Spectrophotometry, Ultraviolet
D009211:Myoglobin; D016650:Fluorescein-5-isothiocyanate