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.
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| 3 | 488 | 660/20 | 492 | 665 | (in buffer) 5 | flow cytometry |
Invitrogen NovaFluor Blue 660 dyes are available with two different levels of brightness—NovaFluor Blue 660-40S and NovaFluor Blue 660-120S—that allow differential matching of the dye brightness to the antigen density or abundance without panel redesign. NovaFluor Blue 660-120S dye is the brighter dye. NovaFluor Blue 660-120S dye can be used to replace PE-tandem dyes, such as PE-Cy5 and PE/Fire™ 640, using the 488 nm blue laser line for excitation. Because PE-tandem dyes are cross-excited by both blue and yellow-green lasers, the replacement of these dyes with NovaFluor Blue 660-120S dye can allow detection of an additional marker labeled with, for example, NovaFluor Yellow 660 dye.
The macromolecule-based NovaFluor Blue 660-120S 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.
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.
- Narrow emission spectrum and minimal cross-laser excitation for better resolution
- Higher intensity (~3 times brighter than NovaFluor Blue 660-40S dye) allows it to be paired with medium-to-low abundance antigens
- Can serve as a replacement for PE-tandem dyes using the blue laser, with reduced cross-laser excitation and fluorescent spillover
- Brighter replacement for PerCP-Cy5.5
- Single PE-tandem dyes can be replaced by the NovaFluor Blue 660-120S and NovaFluor Yellow 660 dyes to allow detection of an additional marker in a flow cytometry panel
- NovaFluor Blue 660-40S and NovaFluor Blue 660-120S can be used together and unmixed using a spectral flow cytometer
- Produces highly stable fluorescence and is compatible with all buffers tested
Click image to enlarge
Spectral signature of NovaFluor Blue 660-120S dye. Normal human peripheral blood cells stained with CD4 Monoclonal Antibody (SK3 (SK-3)), NovaFluor Blue 660-120S were used for analysis. Data was acquired on a 5-laser Cytek Aurora system.
Staining of human peripheral blood cells with NovaFluor Blue 660-120S conjugated antibody. Normal human peripheral blood cells (PBMCs) were unstained (left) or stained with CD19 Monoclonal Antibody, NovaFluor Blue 660-120S (right). Data was acquired in the B7 channel on a 5-laser Cytek Aurora system, and singlet cells 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
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.
Cy™ is a trademark or registered trademark of GE Healthcare.
Not for resale. Super Bright Polymer Dyes are sold under license from Becton, Dickinson and Company.
Brilliant Ultra Violet™ and Brilliant Violet™ are trademarks or registered trademarks of Becton, Dickinson and Company or its affiliates, and are used under license.
For Research Use Only. Not for use in diagnostic procedures.
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