A Bright Fluorochrome Suitable for Low to Moderate Antigen Density Markers
BD Horizon Brilliant™ Violet 786 (BV786) is a tandem fluorochrome consisting of a BV421 donor with an excitation maximum of 407 nm and an acceptor dye with an emission maximum of 786 nm. As part of the BD Horizon Brilliant™ Violet Family of Dyes, BV786 supports flow cytometry applications, including:
- Detection of low to moderately expressed markers
- Conventional and spectral flow cytometry
- Surface and intracellular staining workflows
BV786 is a violet laser-excited fluorochrome (405 nm) with peak emission at 786 nm
| Format | Ex Max | Em Max | Spectral | Conventional | Relative Brightness | Spillover* (1 = low, 4 = high) | Alternative To |
|---|---|---|---|---|---|---|---|
| BV7860 | 407 nm | 786 nm | Yes | Yes | 2 | BV785**, Qdot™ 800, StarBright™ Violet 790 |
*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.
** BV785 is sold by BioLegend
BD Horizon™ BV786-Conjugated Primary Antibodies Clearly Detect Moderate-Expression Markers
Human whole blood was treated with BD Pharm Lyse™ Lysing Buffer to lyse erythrocytes. Leukocytes were washed, preincubated with BD Pharmingen™ Human BD Fc Block™ and stained with PE Mouse Anti-Human CD56 (B159) Antibodies (top) or BD Horizon™ BUV395 Mouse Anti-Human CD3 (UCHT1) Antibodies (bottom), along with BD Horizon™ BV786 Rat Anti-Human CD159a (DD11-740.rMAb) Antibodies. Samples were acquired on a 5-laser Flow Cytometer System and analyzed with FlowJo™ Software.
Multicolor Flow Cytometry
BV786 clearly stains CD123 in a spectral flow cytometry panel
In a 16-color myeloid panel, BV786 was assigned to CD123 (9F5), a critical marker for plasmacytoid dendritic cell analysis. Below is the gating strategy used to analyze myeloid cells from human whole blood, including analysis of activation markers CD69 and CD40.
Myeloid cells were gated on single cells from a lysed whole blood sample by excluding CD3+ T cells and CD19+ B cells. CD303+ CD123+ Plasmacytoid Dendritic Cells (pDCs), CD123+ CD303- Basophils and CD303-CD123- cells were visualized. HLA-DR+ CD11c+ conventional DCs were identified within the SSC Low CD14- Lymphocyte population and further characterized using CD141 and CD1c. Classical, intermediate, and non classical monocyte populations were identified by assessing CD86, CD64, CCR2, HLA-DR, CD16 and CD14 expression within the SSC High population. The expression of activation markers CD69 and CD40 were assessed in pDCs, CD1c+ DCs, Classical Monocytes and CD3+CD19+ Lineage cells. Samples were acquired on a 5-laser BD FACSDiscover A8 Cell Analyzer. Data analysis was performed using FlowJo Software.
FAQs
- What laser and filter set should I use with BV786?
BV786 is excited by the violet laser (405 nm) and has emission maximum at 786 nm. Use a 780/60 nm or similar far-red bandpass filter for detection. Confirm the fluorochrome’s compatibility with your cytometer's laser and detector configuration using the BD® Spectrum Viewer.
- Is BV786 a tandem dye and what does that mean for my workflow?
Yes. BV786 is a tandem composed of BV421 (donor) and an acceptor dye emitting at 786 nm. Like other tandem dyes, batch-specific compensation controls are recommended.
- What should I take into consideration when using BV786 in a multicolor panel?
BV786 may show excitation by UV or red lasers, as indicated by its excitation spectrum, and may contribute residual signal into the BV421 detector. Use the BD® Spectrum Viewer to assess potential spillover and identify fluorochrome assignments appropriate for your panel.
- Should I use BD Horizon™ Brilliant Stain Buffer when using BV786 along with other BD Horizon Brilliant™ Dyes?
Yes. It’s recommended to use BSB when using two or more BD Horizon Brilliant™ Dyes. Dye-dye interactions between the fluorochromes may produce artifacts.
For Research Use Only. Not for use in diagnostic or therapeutic procedures. Not for resale.
QDot is a trademark of Thermo Fisher Scientific. StarBright is a trademark of Bio-Rad Laboratories. BioLegend is the registered trademark of BioLegend, Inc.