Thermo Scientific DyLight Long Stokes Shift Specialty Dyes have fluorescence emission from green to far red with a single excitation peak that matches the 488 laser line and very large Stokes shifts that inhibit quenching.
DyLight Long Stokes Shift Specialty Dyes are a family of labeling agents that provide bright fluorescent detection for multiplexed imaging. The dyes share a common excitation range, while their emission properties extend from the green to the far red wavelengths. The dyes are based on a core coumarin structure that is modified to emit at different wavelengths. Each dye contains an amine-reactive NHS ester for rapid modification of antibodies, proteins, peptides or other biomolecules through amide bond formation. DyLight Long Stokes Shift Dyes can be used individually or combined with DyLight 488 for multiplexed analysis. The excitation and emission peaks are sufficiently separated so as to minimize dye-dye quenching in assays.
- Single excitation wavelength – DyLight Long Stokes Shift Dyes all can be excited together at 488 nm using an Argon laser
- Image multiple targets simultaneously – the four Long Stokes Shift Dyes can be used with DyLight 488 for multiplex fluorescence applications
- NHS ester reactive group – allows immediate labeling of antibodies, proteins, peptides and other amine-containing molecules through amide bond formation
- Multiple solubility options – choose from hydrophilic to hydrophobic dyes to optimize the right dye label for the best performance in an application
- Large Stokes shift inhibits quenching – minimal overlap between excitation and emission peaks inhibits dye-dye quenching effects
- Use in FRET-based assays – the extremely broad separation between excitation and emission peaks allow for use of the Long Stokes Shift Dyes in energy transfer applications where the acceptor dye is spectrally far removed from the excitation wavelength.
Criteria to consider when choosing a DyLight Long Stokes Shift Specialty Dye:
- Excitation and emission wavelengths – all dyes share a common excitation wavelength range with options for emission from the green to the far red wavelengths.
- Water solubility – DyLight Long Stokes Shift Dyes can be initially solubilized in methanol, ethanol, DMF or DMSO with transfer into an aqueous buffer medium for labeling biomolecules
- Quenchers and FRET pairs
- Conjugation to silica particles
- STED microscopy
- Multicolor microscopy
- 4Pi microscopy
- Dual-color 3D nanoscopy
- Dynamic light scattering
- Graded stokes shift
- Duplex enhanced RT-PCR
- High-resolution optical microscopy
Thermo Scientific DyLight Long Stokes Shift Specialty Dye Characteristics.
||Laser and filter sets
||488 laser source
Cy 2 excitation/rhodamine emission
||488 laser source
Cy 2 excitation/633 emission
||488 laser source
Cy 2 excitation/Cy 5 emission
|† Fluorescent dye name appended with LS to indicate the long Stokes shift of the dye
†† Molar extinction coefficient (M-1 cm-1) at the absorption maximum
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- Herz E. et al. (2009) Large stokes-shift fluorescent silica nanoparticles with enhanced emission over free dye for single excitation multiplexing. Macromol Rapid Commun. 30, 1907-10.
- Schmidt R. et al. (2008) Spherical nanosized focal spot unravels the interior of cells. Nat Methods. 5, 539-44.
- Carroll-Portillo A. et al. (2010) Formation of a mast cell synapse: Fc epsilon RI membrane dynamics upon binding mobile or immobilized ligands on surfaces. J Immunol. 184, 1328-38.
- Staudt T. et al. (2007) 2,2'-thiodiethanol: A new water soluble mounting medium for high resolution optical microscopy. Microsc Res Tech. 70, 1-9.
- Lang M. C. et al. (2007) 4pi microscopy with linear fluorescence excitation. Opt Lett. 32, 259-61
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NHS ester crosslinking chemistry technical guide
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