Next-generation laser-specific flow cytometry research reagents
The family of BD Horizon RealYellow™ and BD Horizon RealBlue™ Reagents is bright, clean and laser specific to help streamline the path to your scientific discoveries.
Driven by next-generation BD dye technology and AI-guided fluorochrome selection to optimize spectral positioning, our new family of reagents is specially engineered to deliver reduced spillover to optimize resolution when used with other fluorochromes—helping to expand your experiments on both conventional and spectral flow cytometers.
Interview with Jochen Lamote, Head of VIB Flow Core
BD Biosciences sat down with Jochen Lamote to discuss his experience as a beta tester of flow cytometry reagents – including how BD Horizon RealBlue™ minimizes spillover and simplifies panel design.
Watch the interview to discover why the team at Vlaams Institute for Biotechnology (VIB) Flow Core would recommend BD Horizon RealBlue™ to their colleagues.
BD Horizon RealBlue™ 545 (RB545) Reagents
RB545 reagents are innovative laser-specific fluorochromes excited primarily by the 488-nm blue laser that offer:
- Minimal cross-laser excitation off the 561-nm yellow-green laser
- Moderate brightness fluorochrome to support the detection of high-expression surface and intracellular markers
Download the BD Horizon RealBlue™ 545 Brochure>
RB545 reagents can be used on spectral cytometers to increase the number of experimental parameters.
Performance
BD Horizon™ RB545 Reagents offer minimal cross-laser excitation off the yellow-green laser
Applications
RB545 reagents can resolve high-expression surface and intracellular markers and are available in a wide array of specificities. RB545 reagents can be used with PE and FITC on spectral flow cytometers to meet your specific panel design needs.
RB545 reagents can help improve resolution in high-parameter spectral flow cytometry panels.
Human PBMCs were stained with two sets of reagents, moderately similar CD4 RB545 and CD127 FITC, and highly similar CD4 FITC and CD127 BB515. Co-expressed markers were selected to demonstrate impact of spectral unmixing with different levels of similarity. Resolution was impacted when reagents were highly similar, shown in bottom set of CD4 FITC and CD127 BB515. Samples were acquired on the BD FACSymphony™ A5 SE Cell Analyzer and spectrally unmixed with FlowJo™ Software.
RB545 reagents show stable performance with lot-to-lot consistency across made-to-stock reagents and BD OptiBuild™ On-Demand Reagents. They also offer proven photostability when exposed to typical lab lighting or dimmed core lab lighting.
BD Horizon RealBlue™ 545 Reagents are compatible with a broad range of fixation and permeabilization systems
| Buffers | Results |
| BD FACS™ Lysing Solution and BD Pharm Lyse™ Lysing Buffer | Compatible |
| CellBlox™ Blocking Buffer | Compatible |
| BD Cytofix™ Fixation Buffer | Stable at least 24 hours |
| 1% PFA | Stable at least 24 hours |
| BD Cytofix/Cytoperm™ Fixation and Permeabilization Solution | Compatible with antibody staining before and after fixation |
| BD FACS™ Permeabilizing Solution II | Compatible with antibody staining before and after fixation |
| BD Phosflow™ Perm III Buffer | Compatible with antibody staining before and after fixation |
| EDTA and Heparin | Compatible |
| BD Horizon™ Brilliant Stain Buffer (BSB) | Compatible |
BD Horizon RealBlue™ 780 (RB780) Reagents—the superior alternative to PE-Cy7
RB780 reagents leverage an innovative laser-specific fluorochrome, excited primarily by the 488 nm blue laser to offer.
- Minimal cross-laser excitation off the 561-nm yellow-green laser
- Bright fluorochrome to support the detection of low-expression surface and intracellular markers
- Reduced monocyte background compared to PE-Cy7
Download the BD Horizon RealBlue™ 780 Reagents Brochure >
RB780 reagents can be used instead of PE-Cy7 on conventional flow cytometers or with PE-Cy7 on spectral flow cytometers
RB780 reagents can be used with PE-Cy7 on appropriately configured conventional and spectral cytometers with both a 488-nm blue and 561-nm yellow-green laser to increase the number of experimental parameters.
Performance
BD Horizon™ RB780 Reagents offer minimal cross-laser excitation off the yellow-green laser and reduced background compared to PE-Cy7
Normalized emission profile of PE-Cy7 and RB780 with RB780 minimal excitation off the 561-nm yellow-green laser.
Human whole blood was stained with BD Horizon™ RB780 Human CD4 (SK3) Reagent, acquired and compensated on a BD FACSymphony™ A5 SE Cell Analyzer.
BD Horizon™ RB780 Reagents offer reduced background compared to PE-Cy7
Applications
RB780 reagents can resolve low-expression surface and intracellular markers and are available in a wide array of specificities. RB780 reagents can be used instead of PE-Cy7 on conventional flow cytometers or with PE-Cy7 on spectral flow cytometers to meet your specific panel design needs.
The BD Horizon™ RB780 Reagents show stable performance with lot-to-lot consistency across made-to-stock reagents and BD OptiBuild™ On-Demand Reagents. They also offer proven photostability when exposed to typical lab lighting or dimmed core lab lighting.
BD Horizon RealBlue™ 780 Reagents are compatible with a broad range of fixation and permeabilization systems
| Buffers | Results |
| BD FACS™ Lysing Solution and BD Pharm Lyse™ Lysing Buffer | Compatible |
| CellBlox™ Blocking Buffer | Compatible |
| BD Cytofix™ Fixation Buffer | Stable at least 24 hours |
| 1% PFA | Stable at least 24 hours |
| BD Cytofix/Cytoperm™ Fixation and permeabilization Solution | Compatible with antibody staining before and after fixation |
| BD FACS™ Permeabilizing Solution II | Compatible with antibody staining before and after fixation |
| BD Phosflow™ Perm III Buffer | Compatible with antibody staining before and after fixation |
| EDTA and Heparin | Compatible |
| BD Horizon™ Brilliant Stain Buffer (BSB) | Compatible |
Introducing
BD Horizon RealYellow™ 586 (RY586) Reagents
BD Horizon RealYellow™ 586 (RY586) Reagents are now available as the first in the rapidly growing family of innovative, laser-specific fluorochromes. They are excited primarily by the 561-nm yellow-green laser and offer:
- Minimal cross-laser excitation off the 488-nm blue laser
- Bright fluorescence to detect low-expression surface and intracellular markers
Conventional flow cytometry: Just as bright as PE, but with significantly less cross-laser excitation, RY586 reagents can be used instead of PE on conventional flow cytometers for more flexible panel design.
Spectral flow cytometry: With a distinct spectral profile from PE, RY586 reagents can be used with PE on spectral flow cytometers to increase parameters for deep scientific insights.
Performance
Click through the slides below to view performance data.
Clean & Bright
BD Horizon RealYellow™ 586 Reagents Offer High Resolution with Minimal Cross-Laser Excitation
BD Horizon RealYellow™ 586 Reagents exhibit minimal cross-laser excitation off the blue laser, resulting in significantly less spillover into blue detectors than PE. At the same time, they offer minimal background and high resolution comparable to PE.
Versatile Applications
BD Horizon RealYellow™ 586 Reagents have versatile applications
RY586 reagents can resolve low-expression surface and intracellular markers and are available in a wide array of specificities. RY586 reagents can be used instead of PE on conventional flow cytometers or with PE on spectral flow cytometers equipped with the 561-nm yellow-green laser to meet your specific panel design needs.
Spectral Flow Cytometry
BD Horizon RealYellow™ Reagents can be used with spectral flow cytometry
RY586 can be used with PE on spectral flow cytometers. Since RY586 provides comparable brightness to PE, it is ideal for detecting low-expression markers. Using RY586 and PE in the same spectral flow cytometry panel enables high-dimensional analysis of more low-expression surface and intracellular markers.
Stable & Consistent
Performance stability of BD Horizon RealYellow™ Reagents RY586 offers stable reagent performance with lot-to-lot consistency across made-to-stock reagents and BD OptiBuild™ On-Demand Reagents as well as proven photostability when exposed to typical lab lighting or dimmed core lab lighting.
RY586 made-to-stock reagents and BD OptiBuild™ On-Demand Reagents show consistent performance between each other and across multiple lots.
Human whole blood was stained with human CD4 (SK3) RY586, followed by lysis with BD FACS™ Lysing Solution. Mouse splenocytes were stained with mouse CD8a 53-6.7. All specificities were run on a BD FACSymphony™ A5 SE Cell Analyzer.
BD Horizon RealYellow™ Reagents are compatible with a variety of common fixation and permeabilization systems, including:
- BD Cytofix/Cytoperm™ Fixation/Permeabilization Kit
- BD Pharmingen™ Transcription Factor Buffer Set
- BD FACS™ Lysing Solution
- BD Pharm Lyse™ Lysing Buffer
- BD Cytofix™ Fixation Buffer
- BD FACS™ Permeabilizing Solution 2
- BD Phosflow™ Perm III
- BD Horizon™ Brilliant Stain Buffer (BSB)
- 1% PFA
- EDTA and heparin
- CellBlox™ Blocking Buffer
