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.
Still Using PerCP- and PE-Based Tandem Dyes? Time for an Upgrade.
| Format | Spectral | Conventional | Relative Brightness | Spillover (1-low, 4-high) | Alternative To |
|---|---|---|---|---|---|
| Blue Laser Line | |||||
| RB545 | ✓ | 1 | (Unique position) | ||
| RB613 | ✓ | ✓ | 2 | PE-CF594, PE-Dazzle™ 594, BB630-P2 or StarBright™ Blue 615 | |
| RB670 | ✓ | ✓ | 2 | PE-Cy5, BB660-P2 or StarBright™ Blue 675 | |
| RB705 | ✓ | ✓ | 2 | PerCP-Cy5.5, BB700 or StarBright™ Blue 700 | |
| RB744 | ✓ | ✓ | 1 | BB755-P | |
| RB780 | ✓ | ✓ | 1 | PE-Cy7 or BB790-P | |
| Yellow-Green Laser Line | |||||
| RY586 | ✓ | ✓ | 1 | PE | |
| RY610 | ✓ | ✓ | 1 | PE-CF594 or PE-Dazzle™ 594 | |
| RY703 | ✓ | ✓ | 2 | PE-Cy5.5 or StarBright™ Yellow 720 | |
| RY743 | ✓ | ✓ | 2 | PE/Fire™ 744 | |
| RY775 | ✓ | ✓ | 2 | PE-Cy7 or StarBright™ Yellow 800 | |
Note: PE and PE tandem dyes are excited by both the blue and yellow-green lasers. Be sure to check your instrument configuration
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™ 670 (RB670)
Reagents—A superior alternative to PE-Cy5
RB670 reagents leverage an innovative laser-specific fluorochrome, excited primarily by the 488-nm blue laser to offer:
- Significantly reduced cross-laser excitation off the 561-nm yellow-green laser
- A bright fluorochrome to support the detection of low-expression surface and intracellular markers
- Improved spillover compared to PE-Cy5, NovaFluor™ Blue 660 and StarBright™ Blue 675
- Reduced monocyte background compared to PE-Cy5
- Compatibility with conventional (with the appropriate detector) and spectral flow cytometers
RB670 reagents can be used in place of or along with PE-Cy5 on appropriately configured cytometers
Increase panel design flexibility with RB670 reagents, designed to reduce cross-laser excitation and available in hundreds of antibody specificities.
Performance
BD Horizon RealBlue™ 670 Reagents are a cleaner alternative to PE-Cy5, StarBright™ Blue 675 and NovaFluor™ Blue 660
Normalized emission profile of RB670, PE-Cy5, StarBright™ Blue 675 and NovaFluor™ Blue 660-120s fluorochromes acquired on a BD FACSymphony™ A5 SE Cell Analyzer.
RB670 has minimal monocyte background.
Applications
RB670 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™ RB670 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™ RB670 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 2 | 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.
BD FACSLyric™ Flow Cytometer with the BD FACSuite™ Clinical and BD FACSuite™ applications is an in vitro diagnostic medical device bearing a CE mark.