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.
When building assays, we are presented with hundreds of fluorochrome options and their different properties can make selection feel overwhelming. Check out our Fluorochrome Performance Chart with a handy cheat-sheet to help you prioritise clean fluorochromes and simplify your panel design.
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 Buffer III | Compatible with antibody staining before and after fixation |
| EDTA and Heparin | Compatible |
| BD Horizon™ Brilliant Stain Buffer (BSB) | Compatible |
BD Horizon RealBlue™ 613 (RB613) Reagents
A bright new option for high-parameter and spectral flow cytometry panels
RB613 reagents leverage an innovative laser-specific fluorochrome, excited primarily by the 488-nm blue laser to offer:
- A bright fluorochrome to support the detection of low-expression surface and intracellular markers
- Significantly reduced cross-laser excitation off the 561-nm yellow-green laser
- Compatibility with conventional and spectral flow cytometers with the appropriate detectors
Download the BD Horizon RealBlue™ 613 and BD Horizon RealYellow ™ 610>
RB613 reagents can be used in higher parameter flow cytometry panels to increase biological resolution
RB613 reagents increase panel design flexibility off the 488-nm blue laser, offering significantly reduced cross-laser excitation in hundreds of antibody specificities.
Performance
BD Horizon RealBlue™ 613 Reagents are the best choice in this position for flow cytometers with a 488-nm blue laser
Normalized emission profile of RB613 and PE-CF594 fluorochromes.
RB613 shows reduced spillover spread into the corresponding yellow-green detectors.
Applications
RB613 reagents can clearly resolve low-expression surface and intracellular markers and are available in a wide array of specificities.
BD Horizon RealYellow™ and BD Horizon RealBlue™ Reagents resolve well when run together in a spectral flow cytometry panel.
The BD Horizon™ RB613 Reagents show stable performance with lot-to-lot consistency across made-to-stock reagents and BD OptiBuild™ On-Demand Reagents. They also offer photostability when exposed to typical lab lighting or dimmed core lab lighting.
BD Horizon RealBlue™ RB613 Regents are compatible with a broad range of fixation and permeabilization systems
| Buffers | Results |
| BD FACS™ Lysing Solution and BD Pharm Lyse™ Lysing 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 Buffer III | Compatible with antibody staining before and after fixation |
| EDTA and Heparin | Compatible |
| BD Horizon™ Brilliant Stain Buffer (BSB) | Compatible |
BD Horizon RealBlue™ 705 (RB705) Reagents
The superior alternative to PerCP-Cy5.5 and BD Horizon Brilliant™ Blue 700 (BB700) Reagents
RB705 reagents leverage an innovative laser-specific fluorochrome, excited primarily by the 488-nm blue laser to offer:
- A bright fluorochrome to support the detection of low-expression surface and intracellular markers
- Minimal cross-laser excitation off the 561-nm yellow-green laser
- Reduced background compared to BB700 with no additional wash recommended
Download the BD Horizon RealBlue™ 705 Brochure>
RB705 reagents can be used instead of PerCP-Cy5.5 or BB700 on conventional flow cytometers or with PerCP-Cy5.5 or BB700 on spectral flow cytometers
RB705 reagents increase panel design flexibility off the 488-nm blue laser, offering minimal cross-laser excitation in hundreds of antibody specificities.
Performance
BD Horizon RealBlue™ 705 Reagents provide minimal cross-laser excitation off the 561-nm yellow-green laser
Normalized emission profile of RB705, BB700 and PerCP-Cy5.5 fluorochromes.
Compared to PerCP-Cy5.5 and BB700, RB705 shows lower spillover spread into the corresponding yellow-green, red, UV and violet detectors.
Applications
RB705 reagents can clearly resolve low-expression surface and intracellular markers and are available in a wide array of specificities. RB705 reagents can be used instead of PerCP-Cy5.5 or BB700 on conventional flow cytometers or with PerCP-Cy5.5 or BB700 on spectral flow cytometers to meet your specific panel design needs.
RB705 is a bright fluorochrome that works well to resolve low-antigen expression surface markers such as TIM-3.
BD Horizon RealYellow™ and BD Horizon RealBlue™ Reagents resolve well when run together in a spectral flow cytometry panel.
The BD Horizon™ RB705 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™ 705 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 Buffer III | Compatible with antibody staining before and after fixation |
| EDTA and Heparin | Compatible |
| BD Horizon™ Brilliant Stain Buffer (BSB) | Compatible |
BD Horizon RealBlue™ 744 (RB744) Reagents
A bright new option for high-parameter and spectral flow cytometry panels
RB744 reagents leverage an innovative laser-specific fluorochrome, excited primarily by the 488-nm blue laser to offer:
- A bright fluorochrome to support the detection of low-expression surface and intracellular markers
- Minimal cross-laser excitation off the 561-nm yellow-green laser
- Compatibility with appropriately configured* BD FACSymphony™ Analyzers and spectral flow cytometers
*Filter centered near 750 nm, e.g., 750/30
Download the BD Horizon RealBlue™ 744 Reagent brochure >
RB744 reagents can be used in higher parameter flow cytometry panels to increase biological resolution
RB744 reagents increase panel design flexibility off the 488-nm blue laser, offering minimal cross-laser excitation in hundreds of antibody specificities.
Performance
BD Horizon RealBlue™ 744 Reagents provide minimal cross-laser excitation off the 561-nm yellow-green laser
Normalized emission profile of RB744 fluorochromes
RB744 shows minimal spillover spread into the corresponding yellow-green, red, UV and violet detectors.
Applications
RB744 reagents can clearly resolve low-expression surface and intracellular markers and are available in a wide array of specificities.
BD Horizon™ RB744 Reagent resolves CD25 well
BD Horizon RealYellow™ and BD Horizon RealBlue™ Reagents resolve well when run together in a spectral flow cytometry panel.
The BD Horizon™ RB744 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™ 744 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 Buffer III | 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 Buffer III | 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
BD Horizon RealYellow™ 610 (RY610) Reagents
A bright new option for high-parameter and spectral flow cytometry panels
RY610 reagents leverage an innovative laser-specific fluorochrome, excited primarily by the 561-nm yellow-green laser to offer:
- A bright fluorochrome to support the detection of markers across a variety of expression levels
- A clean alternative to BD Horizon™ PE-CF594 and PE-Dazzle™ 594 Reagents with minimal cross-laser excitation off the 488-nm blue laser
- Compatibility with conventional and spectral flow cytometers with the appropriate detectors
Download the BD Horizon RealBlue™ 613 and BD Horizon RealYellow ™ 610>
RY610 reagents can be used in higher parameter flow cytometry panels to increase biological resolution
RY610 reagents, available in hundreds of antibody specificities, provide improved spillover compared to PE-CF594 on instruments with a 561-nm yellow-green laser for additional panel design flexibility in high parameter and spectral flow cytometry.
Performance
BD Horizon RealYellow™ 610 Reagents have a clean emission profile with minimal spillover for flow cytometers with a 561-nm yellow-green laser
Normalized emission profile of RY610 and PE-CF594 fluorochromes.
RY610 shows reduced spillover spread into the corresponding blue detectors.
Applications
RY610 reagents demonstrate excellent performance for surface and intracellular markers at a variety of antigen densities and are available in a wide array of specificities.
BD Horizon RealYellow™ and BD Horizon RealBlue™ Reagents resolve well when run together in a spectral flow cytometry panel.
The BD Horizon™ RY610 Reagents show stable performance with lot-to-lot consistency across made-to-stock reagents and BD OptiBuild™ On-Demand Reagents. They also offer photostability when exposed to typical lab lighting or dimmed core lab lighting.
BD Horizon RealYellow™ 610 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 |
| 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 Buffer III | Compatible with antibody staining before and after fixation |
| EDTA and Heparin | Compatible |
| BD Horizon™ Brilliant Stain Buffer (BSB) | Compatible |
BD Horizon RealYellow™ 775 (RY775) Reagents
Superior alternative to PE-Cy7 for high-parameter and spectral flow cytometry panels
RY775 reagents leverage an innovative laser-specific fluorochrome, excited primarily by the 561-nm yellow-green laser to offer:
- A bright fluorochrome to support the detection of low expression surface and intracellular markers
- A cleaner alternative to PE-Cy7 and StarBright™ Yellow 800 (SBY800) with reduced cross-laser excitation
- Reduced nonspecific monocyte binding compared to PE-Cy7
- Compatibility with conventional and spectral flow cytometers with the appropriate detectors
Download the BD Horizon RealYellow™ 775 Reagent brochure >
RY775 reagents can be used in higher parameter flow cytometry panels to increase biological resolution
RY775 reagents provide improved spillover compared to PE-Cy7 on instruments with a 561-nm yellow-green laser for additional panel design flexibility in high-parameter and spectral flow cytometry.
Performance
BD Horizon RealYellow™ 775 Reagents are very bright with reduced spillover and monocyte background
Normalized emission profile of RY775, PE-Cy7 and StarBright™ Yellow 800
RY775 shows reduced spillover spread into the corresponding blue detectors.
RY775 provides improved nonspecific binding.
Applications
RY775 reagents demonstrate excellent performance for surface and intracellular markers at low antigen densities and are available in a wide array of specificities.
BD Horizon RealYellow™ and BD Horizon RealBlue™ Reagents resolve well when run together in a spectral flow cytometry panel.
The BD Horizon™ RY775 Reagents show stable performance with lot-to-lot consistency across made-to-stock reagents and BD OptiBuild™ On-Demand Reagents. They also offer photostability when exposed to typical lab lighting or dimmed core lab lighting.
BD Horizon RealYellow™ 775 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 |
| 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 Buffer III | Compatible with antibody staining before and after fixation |
| EDTA and Heparin | Compatible |
| BD Horizon™ Brilliant Stain Buffer (BSB) | Compatible |
BD FACSLyric™ Flow Cytometer with the BD FACSuite™ Clinical and BD FACSuite™ applications is an in vitro diagnostic medical device bearing a CE mark.
All other products mentioned above are for Research Use Only. Not for use in diagnostic or therapeutic procedures.
