NovaFluor Blue 725 excitation shown as dotted line and emission shown as solid red histogram
2488720/24493726(in buffer) 3flow 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.

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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
Experimental data using NovaFluor Blue 725 dye

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

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.

NovaFluor Blue 725 conjugated antibody staining performance with lymphocytes from human peripheral blood
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.