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Qdot 605 probe is an excellent red-fluorescent label with outstanding brightness and photostability for imaging and flow cytometry applications. With an emission maximum of ~605 nm, Qdot 605 probe can be excited using 350, 488, 561, or 594 nm laser lines or standard TRITC filter. Optimum imaging results with brighter signals are achieved with a Qdot 605 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 |
35 | 350, 488, 561, 594 | RFP | UV | 605 |
Stain index | 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.
Flow cytometric analysis of whole blood cells using streptavidin conjugates of Qdot probes.
Ammonium chloride lysed whole blood cells were stained with mouse anti human CD3 biotin, washed, stained with Qdot 605 streptavidin conjugate, washed, stained with mouse anti human CD56 biotin, washed, and stained with Qdot 705 streptavidin conjugate, washed, resuspended, and analyzed by flow cytometry using 405 nm excitation and detected using 605⁄20 nm and 710⁄20 nm emission bandpass filters. The dot plot shows Qdot 705 streptavidin conjugate on the Y-axis and Qdot 605 streptavidin conjugate on the X-axis.
We’ve developed functionalized versions of the Qdot 605 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.