Spectraviewer image of NovaFluor Red 725 dye

NovaFluor dyes, including NovaFluor Red 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 Red 725 dye has a unique emission spectrum when excited by the 637nm 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 Red 725 dye should generally be paired with moderately to lowly expressed antigens to minimize spectral spillover in complex multicolor panels. The macromolecule-based NovaFluor Red 725 dye produces highly stable fluorescence, and stained samples retain their fluorescence intensity and spectral signature when stored at 4°C.

Benefits of NovaFluor Red 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 replace dim dyes, such as Alexa Fluor 700 in conventional flow cytometry
  • Produces stable fluorescence and is compatible with all buffers tested

Use NovaFluor dyes with CellBlox Blocking Buffer to reduce background and to block non-specific binding of NovaFluor labels, PE and APC tandems observed with macrophages and monocytes.

NovaFluor Red 725 dye dashboard


Initial brightness
 
 
 
 
 
The NovaFluor Red 725 dye, excited by the red laser is optimized for use for cell surface applications. Can be used for spectral and conventional flow cytometry applications. 
  
  

Photostability in buffer
 
 
 
 
 
  
 637720/30636725
 
Laser line
Common filter set
Excitation max
Emission max		 

NovaFluor Red 725 dye products

We offer NovaFluor dyes conjugated to primary antibodies for use in flow cytometry, as well as NovaFluor Antibody Conjugation Kits, NovaFluor CD4 Label Characterization Kits, and custom conjugation services.

Learn more here:

Experimental data using NovaFluor Red 725 dye products

Figure 1. Absorption and fluorescence emission spectra of NovaFluor Red 725 dye.

Figure 2. Spectral signature of NovaFluor Red 725 dye. Normal human peripheral blood cells stained with anti-CD4 antibodies (clone SK3) conjugated to NovaFluor Red 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 CD3-stained and CD45R-stained vs CD3-stained

Figure 3. C57BL/6 mouse splenocytes were stained with CD3e monoclonal antibody, eFluor 450 dye (Cat. No. 48-0037-42) only (left) or along with CD45R monoclonal antibody, NovaFluor Red 725 dye (Cat. No. H029T03R05) (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 PBMCs, unstained vs CD19-stained and CD4-stained vs CD19-stained

Figure 4. Normal human peripheral blood cells were stained with CD19 monoclonal antibody, eFluor 450 dye (Cat. No. 48-0199-42) only (left) or with CD4 monoclonal antibody, NovaFluor Red 725 dye (Cat. No. H001T03R05) (right). Cells in the lymphocyte gate were used for analysis.