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Qdot 525 probe is an excellent green-fluorescent label with outstanding brightness and photostability for imaging and flow cytometry applications. With an emission maximum of ~525 nm, Qdot 525 probe can be excited using a 488 nm laser line or standard FITC filter. Optimum imaging results with brighter signals are achieved with a Qdot 525 filter or light cube.
Made from nanocrystals of semiconductor material, Qdot probes are broadly compatible with standard fluorescence techniques, but there are some novel aspects to their use that can be optimized to obtain the best possible results. In general they have an extremely broad excitation range and narrow emission with a large Stokes shift. Optimal brightness is generally achieved using a specific Qdot filter.
Organic fluorophores and Qdot probes can be readily combined in multiplexed flow cytometry or imaging studies. Longer-wavelength Qdot probes are also ideal for multiplexed western blots.
Initial brightness | For best results use a Qdot-optimized filter and PBS or a mounting medium without antifade.
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Photostability in buffer |
350, 488 | FITC | UV | 525 | ||
Laser line | Common filter set | Excitation max | Emission max |
The exceptional brightness and photostability of Qdot probes make it easier to detect low-abundance targets. With narrow and symmetric emission spectra, Qdot probes are also ideal for multiplexed fluorescence detection using a combination of Qdot probes and organic dyes.
We’ve developed functionalized versions of the Qdot 525 label for the preparation of custom conjugates of proteins and other biomolecules. The first—a carboxyl-derivatized form—can be coupled to amine groups of proteins and modified oligonucleotides. The second—an amine-derivatized PEG form—can be coupled to isothiocyanates and succinimidyl esters (or with native carboxylic acids using water-soluble carbodiimides).
Amine | Aldehyde-, ketone, carboxylate or phosphate |
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Qdot Probes are made from nanocrystals of semiconductor material and are broadly compatible with standard fluorescence techniques, but there are some novel aspects to their use that can be optimized to obtain the best possible results.