BD Horizon Brilliant™ Violet 711 Reagents

A Bright Fluorochrome to Expand Multicolor Panel Design

The BD Horizon Brilliant^™ Violet 711 (BV711) Dye is a bright and versatile dye in the BD Horizon Brilliant^™ Violet Family of Dyes. Driven by BD innovation, this tandem fluorochrome consists of a BV421 donor with an excitation maximum (Ex max) of 407 nm and an acceptor dye with an emission maximum (Em max) at 713 nm. BV711 delivers:

Reliable performance across markers with varying expression levels
Compatibility with conventional and spectral flow cytometry
Broad marker availability across the BD antibody portfolio

BV711 is excited by the violet laser (405 nm) with peak emission at 713 nm

Format	Ex _Max	Em _Max	Spectral	Conventional	Relative Brightness	*Spillover (1 = low, 4 = high)**	Alternative To
BV711	407 nm	713 nm	Yes	Yes		3	StarBright^™ Violet 710

*Value may vary based on instrument configuration and settings. Spillover ranking is based on cross-laser excitation on five-laser spectral instruments and does not take into account spillover into adjacent detectors.

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Performance

BD Horizon™ BV711 Offers Reliable Performance for Surface and Intracellular Markers

BV711 resolves CD8 well in human whole blood

Human whole blood was lysed with BD Pharm Lyse™ Lysing Buffer, washed, and then preincubated with BD Pharmingen™ Human BD Fc Block™. Cells were then stained with BD Horizon™ BV711 Mouse Anti-Human CD8 Antibody (SK1). Cells were acquired on a BD LSRFortessa™ X-20 Cell Analyzer System and analyzed using FlowJo™ Software.

BV711 Delivers Clear Resolution of IFN-γ-expressing Populations in Intracellular Staining Workflows

Mouse splenocytes were stimulated with PMA and Ionomycin in the presence of BD GolgiStop^™ Protein Transport Inhibitor. Cells were harvested, washed with BD Pharmingen™ Stain Buffer (FBS), and fixed with BD Cytofix^™Fixation Buffer. Cells were then washed and stained in BD Perm/Wash™ Buffer with PE Rat Anti-Mouse CD4 antibody and BD Horizon BV711 Rat Anti-Mouse IFN-γ Antibody (XMG1.2). Flow cytometric analysis was performed using a BD^® LSR II Flow Cytometer and analyzed using FlowJo^™ Software.

Multicolor Flow Cytometry

BV711 supports reliable performance in a 21-color flow cytometry panel

In a 21-color flow cytometry panel of fresh PBMCs, BV711 was assigned to CD117, an essential and critical marker in identifying the ILC subsets. Below is the gating strategy used to analyze ILC groups in fresh PBMCs and protein expression patterns in the three ILC1 subsets.

A. Cells were gated based on conventional phenotypes, ILC1 (CD117^-CD294^-), ILC2 (CD117⁺/CD294⁺), ILC progenitor cells ILCP/ILC3 (CD117⁺/CD294^-). Samples were acquired on the BD FACSymphony A5 Cell Analyzer. Lineage^-CD3^-CD56^-CD127⁺CD117^-CD294^- ILC1 cells were further gated into three subsets: CD4^-CD8^- (blue), CD4⁺/CD8^- (red) and CD4^-/CD8⁺ (green).

B. Protein expression patterns across the three ILC1 subsets.

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FAQs

What is BV711?
BV711 is a tandem fluorochrome in the BD Horizon Brilliant^™ Violet Dye family and consists of a BV421 donor and acceptor dye. Its excitation maximum is 407 nm and emission maximum is 713 nm. It is excited by a 405 nm violet laser and is compatible with spectral and conventional flow cytometry. It is a bright fluorochrome and an alternative to Bio-Rad StarBright^™ V710.
What is the optimal filter for BV711 on a conventional cytometer?
BV711 is best detected using filters centered near 710 nm such as the 712/20 nm bandpass filter.
How should BV711 be assigned to markers in a multicolor flow cytometry panel?
BV711 is a bright dye that can perform well for markers with low to moderate expression and those expressed over a continuum.
How should BV711 be assigned to manage spectral overlap between dyes?
Review adjacent spectral overlap with neighboring fluorochromes before finalizing the panel (the BD ^® Spectrum Viewer can help you do this). One thing to note is that BV711 has very low adjacent spillover into RV828, making it a favorable pairing for spectral panels. However, there is cross-laser spectral overlap with BUV737, RY703, R718 and APC-R700 on both conventional and spectral instruments. Therefore, use caution when assigning these dyes, as increased spread may reduce biological resolution.
Can BV711 be used for intracellular staining?
Yes. We have validated BV711 for intracellular cytokine staining with the BD Cytofix/Cytoperm™ Fixation/Permeabilization Kit. BV711 can also be used for transcription factor workflows using the BD Pharmingen™ Transcription Factor Buffer Set. BV711 is also compatible with BD Phosflow™ Perm Buffer III.