Seta-670-mono-NHS (K8-1342), a dye that has been recently used in single molecule, homo-FRET measurements showed a remarkably low blinking effect which is an important factor in such measurements:
Wide field image for anti-rabbit IgG antibodies labeled with Seta-670 (K8-1342) deposited on a glass slide (D/P=3.2) obtained using 635 nm excitation (~4 mW) and observed through a 685/35 nm bandpass filter. (A) before and (B) after photobleaching of a single IgG protein/spot (marked by arrow). (C) typical fluorescence intensity trajectory during photobleaching illumination (~12 mW) for a D/P=3.2 conjugate. The selected spot undergoes a three steps irreversible photobleaching. The inserts show the time-resolved photon histograms and calculated fluorescence lifetimes for the corresponding regions of fluorescence intensity trajectory
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References |
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Luchowski R., Matveeva E.G., Gryczynski I., Terpetschnig E.A., Patsenker L., Laczko G., Borejdo J., Gryczynski Z. Single molecule studies of multiple-fluorophore labeled antibodies. Effect of homo-FRET on the number of photons available before photobleaching. Current Pharmaceutical Biotechnology, 9, 411-420 (2008). |
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