Fluorescein isothiocyanate monitoring

Labelling Na,K-ATPase with ATP analogues provides evidence for contribution from charged residues that are widely separated in the sequence of a subunit of Na,K-ATPase. The first indication came from ATP sensitive covalent insertion of fluorescein-isothiocyanate (FITC) into Lys ° in the a subunit [90], The strong fluorescence signal provides a convenient probe for monitoring conformational transitions in the proteins. Site-directed mutagenesis of Lys reduces the activity of... 

T. Konecsni and F. Kilar, Monitoring of the conjugation reaction between human serum transferrin and fluorescein isothiocyanate by capillary electrophoresis. J. ChromatogrA, 1051 (2004) 135-139. 

PR Banks, DM Paquette. Monitoring of a conjugation reaction between fluorescein isothiocyanate and myoglobin by capillary zone electrophoresis. J Chromatography 693 145-154, 1995. 

Receptor-binding activity of the protein-folate conjugate can be determined by competitive displacement of a known quantity of [3H]-labeled folic acid (available from NEN Life Science Products, Boston, MA) from receptor-expressing cells. Alternatively, if the protein can be labeled with a fluorescent probe (e.g., fluorescein isothiocyanate), its uptake by folate receptor-bearing cells can be monitored by fluorescence microscopy (8). 

The function of the device containing 2-mm long monolith was first tested on capture of fluorescein isothiocyanate (FITC)-labeled lysozyme from its mixture with cytochrome c. The capture was monitored via an increase in fluorescence of the monolith. The ability of the monolith to selectively capture lysozyme from the mixture was validated by MALDI-TOF mass spectrometry. Since the Cibacron-blue-3G-A ligand has a strong affinity for albumins, the ultimate function was successfully demonstrated with capture of albumin from human cerebrospinal fluid. ... 

Effects produced by microinjected lamin antibodies were monitored by immu-nocytochemistry and electron microscopy. For immunocytochemistry, after different incubation times following microinjection, cells were fixed in methanol (-20°C, 10 min) and acetone (-20°C, 1 min), and then air dried. Microinjected cells were identified by incubating the cover sUps (10 min) with antiguinea pig secondary antibodies conjugated to fluorescein isothiocyanate (FITC Dianova, Hamburg, Germany). 

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