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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| 4 | 405 | 450/50 | 406 | 423 | (in buffer) 5 | flow cytometry |
Brilliant Violet dyes, including Brilliant Violet 421 (BV421), are a polymer dye-based technology and compatible with both spectral flow cytometry, as well as traditional flow cytometry. The violet laser has unique channels far from heavily occupied detectors allowing for larger panels. BV421 is excited by the 405 nm violet laser and emits at 421 nm. This dye has a medium-to-high relative brightness and can be used for cell surface, intracellular, and nuclear staining.
When using two or more Super Bright, Brilliant Violet, Brilliant Ultra Violet™, or other polymer dye- conjugated antibodies in a staining panel, we recommend that you use Super Bright Complete Staining Buffer or Brilliant Stain Buffer to minimize any non- specific polymer interactions.
We offer BV421 dye conjugated to primary antibodies for use in flow cytometry. BV421 can also be used for spectral flow cytometry applications.
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Figure 1. Excitation and emission of Brilliant Violet™ 421 dye (BV421).
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Figure 2. Spectral signature of BV421 dye. Data acquired on a 5-laser Cytek Aurora and normal human peripheral blood cells stained with clone RPA-T8 (hCD8) conjugated to BV421 dye were used for analysis.
Figure 3. Normal human peripheral blood cells were unstimulated (left) or stimulated for 5 hours with the Cell Stimulation Cocktail (plus protein transport inhibitors) (right). Cells were then stained intracellularly, using the Intracellular Fixation & Permeabilization Buffer Set and protocol, with CD4 Monoclonal Antibody, FITC and TNF alpha Monoclonal Antibody, Brilliant Violet™ 421 dye. Viable cells in the lymphocyte gate were used for analysis, as determined by Fixable Viability Dye eFluor 780 dye.
Figure 4.C57BL/6 mouse splenocytes were stained with CD3e Monoclonal Antibody, FITC and 0.06 µg of Rat IgG2a kappa Isotype Control, Brilliant Violet™ 421 dye (left) or 0.06 µg of CD4 Monoclonal Antibody, Brilliant Violet™ 421 dye (right). Viable cells in the lymphocyte gate were used for analysis, as determined by 7-AAD Viability Staining Solution.
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.