Graph of NovaFluor Blue 725 dye excitation and emission
3488710/50492725(in buffer) 3flow cytometry

NovaFluor dyes, including NovaFluor Blue 725 dye, 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.

NovaFluor Blue 725 dye has a unique emission spectrum when excited by the 488nm 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.

Benefits of NovaFluor Blue 725 dye

  • 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

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.

NovaFluor Blue 725 primary antibodies

Graph of fluorescence intensity vs channel for NovaFluor Blue 725 dye

Figure 1. 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.

2-panel scatter plots of mouse splenocytes, unstained vs CD3e-stained and CD8a-stained vs CD3e-stained

Figure 2. NovaFluor Blue 725 conjugated antibody staining performance with mouse C57BL/6 mouse splenocytes were stained with CD3е Monoclonal Antibody, eFluor 450 only (left) or along with CD8a Monoclonal Antibody, NovaFluor Blue 725 (right). Viable cells were used for analysis, as determined by LIVE/DEAD Fixable Blue Dead Cell Stain Kit (Cat. No. L34962).

2-panel scatter plots of human lymphocytes, unstained vs CD19-stained and CD3-stained vs CD19-stained

Figure 3. NovaFluor Blue 725 conjugated antibody staining performance with human PBMCs. 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.

Additional resources

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