Figure 1. Spectral signature of Alexa Fluor 660.
Alexa Fluor 660 has a unique signature optimized for flow cytometers with spectral capabilities. This fluorophore can be used for cell surface, intracellular, and intranuclear antigens. It should be used in complex multicolor panels to fit between fluorophores Alexa Fluor 647 and Alexa Fluor 700.
Alexa Fluor 660 dye dashboard
|The red laser–excitable Alexa Fluor 660 dye is optimized for use in spectral flow cytometry applications.|
Photostability in buffer
Figure 2. CD4-Alexa Fluor 660 example. C57BL/6 mouse splenocytes were stained with CD3 Monoclonal Antibody, Alexa Fluor 700 (Cat. No. 56-0033-82), CD19 Monoclonal Antibody, APC (Cat. No. 17-0193-82), and CD25 Monoclonal Antibody, Alexa Fluor 647 (Cat. No. 51-0251-82) and 0.125 µg of CD4 Monoclonal Antibody, Alexa Fluor 660. Cells in the lymphocyte gate were used for analysis and gated on CD3 positive/CD19 negative cells (left). CD3 positive cells were further plotted as CD4 versus CD25 to determine expression of CD25 (right). This data was collected on a 5-laser Cytek Aurora full spectral cytometer. As shown here, Alexa Fluor 660 is compatible with Alexa Fluor 647 and Alexa Fluor 700 when proper panel design principles are applied.
Figure 3. Foxp3-Alexa Fluor 660 staining. BALB/c mouse splenocytes were surface-stained with CD25 Monoclonal Antibody, Alexa Fluor 488 (Cat. No. 53-0251-82). Cells were then stained intracellularly, using the Foxp3/Transcription Factor Staining Buffer Set (Cat. No. 00-5523-00) and protocol, with 0.25 µg of Rat IgG2a kappa Isotype Control, Alexa Fluor 660 (Cat. No. 606-4321-81) (left) or 0.25 µg of FOXP3 Monoclonal Antibody, Alexa Fluor 660 (right). Cells in the lymphocyte gate were used for analysis. This data was collected on a 5-laser Cytek Aurora full spectral cytometer.
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