Staining of mouse splenocytes with NovaFluor Blue 725 conjugated antibody. C57BL/6 mouse splenocytes were stained with CD3 Monoclonal Antibody, eFluor 450 only (left) or with CD8a Monoclonal Antibody, NovaFluor Blue 725 (right). Viable cells were used for analysis, as determined by LIVE/DEAD Fixable Violet Dead Cell Stain Kit, for 405 nm excitation.
 |  |  |  |  |  |  |
| 2 | 488 | 720/24 | 493 | 726 | (in buffer) 3 | flow cytometry |
Invitrogen NovaFluor Blue 725 dye has a unique emission spectrum when excited by the 488 nm laser, which fits in a spectral space that is currently not accessible by other spectral dyes. Adding spectrally unique dyes may allow for building out larger panels with minimal loss of resolution.
NovaFluor Blue 725 dye should generally be paired with moderately to highly expressed antigens in multicolor panels. The macromolecule-based NovaFluor Blue 725 dye produces highly stable fluorescence, and stained samples retain their fluorescence intensity and spectral signature when stored at 4°C. Use NovaFluor dyes with CellBlox Plus Blocking Buffer to reduce background and to block non-specific binding of NovaFluor labels, PE and APC tandems observed with macrophages and monocytes.
- Narrow emission spectrum and minimal cross-laser excitation for better resolution
- Should be paired with moderately expressed antigens to minimize spillover
- Spectrally unique and can be used to fill unused detectors in conventional flow cytometry
- Produces stable fluorescence and is compatible with all buffers tested
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Spectral signature of NovaFluor Blue 725 dye. Normal human peripheral blood cells stained with anti-CD4 antibodies (clone SK3) conjugated to NovaFluor Blue 725 dye were used for analysis. Data was acquired on a 5-laser Cytek Aurora system.
Staining of human peripheral blood cells with NovaFluor Blue 725 conjugated antibody. Normal human peripheral blood cells were stained with CD19 Monoclonal Antibody, eFluor 450 only (left) or with CD3 Monoclonal Antibody, NovaFluor Blue 725 (right). Cells in the lymphocyte gate were used for analysis.
NovaFluor dyes
NovaFluor dyes are built using Phiton technology and are compatible with both spectral flow cytometry and traditional flow cytometry. NovaFluor dyes exhibit narrow emission spectra and minimal cross-laser excitation, which helps reduce spectral spillover for better marker resolution. In addition, their unique spectral signatures can provide the opportunity to detect additional markers in flow cytometry panels by opening up previously unusable channels.
Additional resources
Flow cytometry protocols handbook
Protocols that fit your needs in flow cytometry ranging from sample preparation to numerous cell stimulation conditions, staining, immunophenotyping, and data analysis strategies.
Fluorophore and reagent selection guide for flow cytometry
A handy reference poster featuring the broad range of our available dyes and labeling reagents.
Imaging protocol handbook
Protocols that fit your needs in imaging ranging from sample and assay preparation to staining, labeling, and data analysis strategies.
Fluorophore and reagent selection guide for cell imaging
A tool for selecting the optimal fluorescent dyes for your Invitrogen EVOS cell imaging systems.
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