Added labels are deposited proximal to the enzyme site, resulting in minimal loss of resolution and significant enhancement of the fluorescent signals. The biotinyl-tyramide can be detected fluorescently with antifluorescein HRP or antifluorescein AP plus streptavidin conjugated with fluorescein or indirectly with a chromogenic substrate. The availability of four fluorophore-conjugated tyramides enables sequential, multianalyte detection protocols or combination of protein and nucleic acid detection on the same slide. But fluorescent methods are also being used to replace standard protein quantitation methods. Molecular Probes provides two such basic systems suitable for HTS applications.
Researchers can now even engineer proteins to contain endogenous fluorogenic signals.
Upon cleavage of the molecule, the fluorophore is released and becomes brightly fluorescent. Despite the multitude of fluorescence detection methods and applications described, it is safe to say that the power of this approach has not been fully exploited. Stay tuned for advances in miniaturization, multiparameter analysis, single-molecule detection, and robotics. Inc. All rights reserved. We welcome your opinion.