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Qdot 655 probe is an excellent far red–fluorescent label with outstanding brightness and photostability for imaging and flow cytometry applications. With an emission maximum of ~655 nm, Qdot 655 probe can be excited using a 350, 488, 595, or 615 nm laser line or standard Cy5 filter. Optimum imaging results with brighter signals are achieved with a Qdot 655 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 |
20 | 350, 488, 595, 615 | Cy5 | UV | 655 |
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.
BPAE cells were fixed and permeabilized using the Image-iT Fixation/Permeabilization Kit and incubated with Anti- ATP Synthase Subunit IF1 followed by GAM-QD655. Cells were labeled with ReadyProbes reagents NucBlue Live and ActinGreen 488, then mounted using ProLong Gold Antifade Reagent. Images were taken using the EVOS FL Auto Imaging System and a 40X CoverSlip corrected Objective.
We’ve developed functionalized versions of the Qdot 655 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. Learn more about Qdot Probes: