BD OptiBuild Reagents

Sample Data

The on-demand nature of BD OptiBuild™ reagents is the result of years of testing and analysis. We can offer robust reagents that provide a valuable alternative to traditional reagents. The sample data below tells a story of the journey to expanding our portfolio of reagents to simplify panel design.

Panel design is critically important to generating high-quality data. Principles of panel design dictate the optimal matching of antigen density with fluorochrome brightness, yet at least one required reagent is unavailable for approximately 20% of panels designed today. Access to the right reagents can improve resolution and data quality, ultimately leading to deeper scientific insights through your research.


Expanding options for multicolor panels

BD OptiBuild reagents provide choice, so that you have multiple options for your markers of interest. BD OptiBuild and BD Horizon™ reagents complement each other so that you can optimize your panel with a reliable reagent in the color you need.

CD21 fluorescence on multiple formats of BD Horizon Brilliant and BD OptiBuild reagents
CD21 fluorescence on multiple formats of BD Horizon Brilliant and BD OptiBuild reagents
Whole blood was stained with BV421 (Cat. No. 562966), BV510 (Cat. No. 740165), BV605 (Cat. No. 740395), BV650 (Cat. No. 740569), BV711 (Cat. No. 563163), BV786 (Cat. No. 740969) or BUV395 (Cat. No. 740288) mouse anti-human CD21 (clone B-Ly4). The fluorescence density plots show CD21 fluorescence and side light-scatter characteristics of viable cells.
CD11b fluorescence on multiple formats of BD Horizon Brilliant and BD OptiBuild reagents
CD11b fluorescence on multiple formats of BD Horizon Brilliant and BD OptiBuild reagents
Whole blood was stained with BV421(Cat 562632), BV480 (Cat 746704), BV510 (Cat 563088), BV605 (Cat 562721), BV650 (Cat 740566), BV711 (Cat 740771), BV786 (Cat 740965), BUV395 (Cat 563839), or BB700 (Cat 742210) CD11b (clone ICRF44). The fluorescence density plots show CD11b fluorescence and side light-scatter characteristics of viable cells. The D# in the lower right of the plots indicate the different donors that were used to assay the reagent performance.
KLRG1 fluorescence on multiple formats of BD Horizon Brilliant and BD OptiBuild reagents
KLRG1 fluorescence on multiple formats of BD Horizon Brilliant and BD OptiBuild reagents
Fresh balb/c splenocytes were stained with BV421(Cat. No. 562897), BV510 (Cat. No. 740156), BV605 (Cat. No. 564013), BV650 (Cat. No. 740553), BV711 (Cat. No. 564014), BV786 (Cat. No. 565477) or BUV395 (Cat. No. 740279) Hamster Anti-Mouse KLRG1 (clone 2F1) antibody and APC or PE Mouse Anti-Mouse NK1.1 (clone PK136). The fluorescence density plots were derived from gated events with the forward and side light-scatter characteristics of viable cells.

BD Horizon™ BB700: A brighter alternative to PerCP-Cy™5.5

The BD OptiBuild™ portfolio now includes BD Horizon Brilliant™ Blue 700. BB700 was developed as a brighter alternative to PerCP-Cy™5.5.

Human CD166 Comparison
Human CD166 Comparison

Whole blood was stained with BB700 (Cat 745746) or PerCP-Cy5.5 (Cat 562131) mouse anti-human CD166 (clone 3A6).

Human CD23 Comparison
Human CD23 Comparison

Whole blood was stained with BB700 (Cat 745756) or PerCP-Cy5.5 (Cat 561166) mouse anti-human CD23 (clone M-L233).

Mouse CD162 Comparison
Mouse CD162 Comparison

Fresh Balb/c splenocytes were stained with BB700 (Cat 742183) or PerCP-Cy5.5 (Cat 564310) rat anti-mouse CD162 (clone 2PH1).

Mouse CD38 Comparison
Mouse CD38 Comparison

Fresh balb/c splenocytes were stained with BB700 (Cat 742132) or PerCP-Cy5.5 (Cat 562770) rat anti-mouse CD38 (clone 90/CD38).


Reagent choice brings flexibility to your research, but it’s important to keep in mind the BD Horizon principles of matching antigen density with fluorochrome brightness to design optimized panels. Depending on your applications and research goals, some dyes might resolve your populations better than others. The Research Applications team is an excellent resource to discuss options and receive guidance on what will be best for your experiments.

CD21 expression on multiple formats of BD Horizon Brilliant and BD OptiBuild reagents
CD21 expression on multiple formats of BD Horizon Brilliant and BD OptiBuild reagents
Whole blood was stained with BV421 (Cat. No. 562966), BV510 (Cat. No. 740165), BV605 (Cat. No. 740395), BV650 (Cat. No. 740569), BV711 (Cat. No. 563163), BV786 (Cat. No. 740969) or BUV395 (Cat. No. 740288) mouse anti-human CD21 (clone B-ly4) and counter-stained with CD19. All formats are able to resolve CD21 expression on B cells. However, depending on the ability of a dye to resolve dim populations, a subset of T cells may be identified expressing CD21.

BD OptiBuild, BD Horizon Brilliant™ comparisons

Although BD OptiBuild reagents differ from the way our traditional reagents are manufactured and quality control tested, these reagents maintain the high performance and quality standards of the BD Horizon Brilliant™ dyes.

Mouse CD25 direct comparison*
Mouse CD25 direct comparison

Fresh balb/c splenocytes were stained with APC Rat Anti-Mouse CD4 (Cat. No. 553051) and either BD Horizon BV421 Rat Anti-Mouse CD25 (clone PC61, Cat. No. 562606), left panel, or the BD OptiBuild equivalent, right panel. The fluorescence density plots were derived from gated events with the forward and side light-scatter characteristics of viable cells.

Mouse CD31 direct comparison*
Mouse CD31 direct comparison

Fresh balb/c bone marrow was stained with APC Rat Anti-Mouse CD45R/B220 (Cat. No. 557683) and either BD Horizon BV421 Rat Anti-Mouse CD31 (clone MEC 13.3) antibody (Cat. No. 562939), left panel, or the BD OptiBuild equivalent, right panel. The fluorescence density plots were derived from gated events with the forward and side light-scatter characteristics of viable cells.

Mouse CD184 direct comparison*
Mouse CD184 direct comparison

Fresh balb/c thymocytes were stained with APC Rat Anti-Mouse CD4 (Cat. No. 553051) and either BD Horizon BV421 Rat Anti-Mouse CD184 (clone 2B11/CXCR4) antibody (Cat. No. 562738), left panel, or the BD OptiBuild equivalent, right panel. The fluorescence density plots were derived from gated events with the forward and side light-scatter characteristics of viable cells.

Human CD28 direct comparison*
Human CD28 direct comparison

Whole blood was stained with either BD Horizon BV421 Mouse Anti-Human CD28 (clone 28.2, Cat. No. 562613), left panel, or the BD OptiBuild equivalent, right panel. The fluorescence density plots show BV421 signal vs side light-scatter characteristics of viable cells.

Human CD14 direct comparison*
Human CD14 direct comparison

Whole blood was stained with either BD Horizon BV421 Mouse Anti-Human CD14 (clone MfP9, Cat. No. 563743), left panel, or the BD OptiBuild equivalent, right panel. The fluorescence density plots show BV421 signal vs side light-scatter characteristics of viable cells.

Human CD33 direct comparison*
Human CD33 direct comparison

Whole blood was stained with either BD Horizon BV421 Mouse Anti-Human CD33 (clone WM53, cat. 562854) left panel, or the BD OptiBuild equivalent right panel. The fluorescence density plots show BV421 signal vs side light-scatter characteristics of viable cells.

Human CD4 BB700 direct comparison*
Human CD4 BB700 direct comparison

Whole blood was stained with either BD Horizon BB700 Mouse Anti-Human CD4 (clone SK3, cat. 566392) left panel, or the BD OptiBuild equivalent right panel. The fluorescence density plots show BB700 signal versus side light-scatter characteristics of viable cells.

Human CD4 BV480 direct comparison*
Human CD4 BV480 Comparison

Whole blood was stained with either BD Horizon BV480 Mouse Anti-Human CD4 (Clone SK3, cat. 566104) left panel, or the BD OptiBuild equivalent right panel. The fluorescence density plots show BV480 signal versus side light-scatter characteristics of viable cells.


BD OptiBuild performance in multicolor panels

BD OptiBuild reagents can be used to redesign or expand upon existing panels that have various combinations of fluorochromes and antibodies. BD OptiBuild reagents have been extensively tested in multicolor cocktails to demonstrate reproducible results compared to traditional reagents.

Human 8-color T-cell panel direct comparison with and without BD OptiBuild reagents
Human 8-color T-cell panel direct comparison with and without BD OptiBuild reagents
Whole blood was stained with BUV496 CD3 (Cat. No. 564810), BUV395 CD4 (Cat. No. 563550 or BD OptiBuild), APC-Cy7 CD8 (Cat. No. 557834), BV421 CD27 (Cat. No. 562513) or BD OptiBuild, APC CD45RA (Cat. No. 550855), PE-CF594 CD197 (Cat. No. 562381), PE CD25 (Cat. No. 555432), and BV786 CD127 (Cat. No. 563324) and then analyzed for various T-cell subsets. Inclusion of BD OptiBuild products does not have an impact on performance in multicolor cocktails.
Optimized multicolor panel designed with BD OptiBuild reagents
Optimized multicolor panel designed with BD OptiBuild reagents
Whole blood was stained with BV510 CD3 (Cat. No. 563109), PerCP-Cy™5.5 CD4 (Cat. No. 560650), BUV395 CD45RA (Cat. No. 740298), BV421 CD197 (Cat. No. 562555), BB515 CD25 (Cat. No. 564467), and BV786 CD127 (Cat. No. 53324) and then analyzed for various T-cell subsets. Flow cytometry was performed using a BD FACSCelesta™ flow cytometer system (Cat. No. 660346).

Lot-to-lot consistency

BD OptiBuild reagents are made on demand, which means that every reagent vial is equivalent to a new batch. We know that consistent performance is of the utmost importance for the reliability of research results, so the manufacturing process was also extensively tested for lot-to-lot consistency.

Overlapping histograms of six lots of Ms CD8a BV421 at three concentrations
Overlapping histograms of six lots of Ms CD8a BV421 at three concentrations
Fluorescence histogram showing mouse CD8a expression identified using six separate lots of BD OptiBuild conjugated antibodies made with the same dye and antibody lots. All six lots were assessed at three different concentrations. Data was derived from gated events with the forward and side light-scatter characteristics of intact splenocytes.


The data above shows multiple conjugation events using the same batches of dye and antibody, demonstrating highly reproducible performance. We also assessed the impact of other potential sources of variability by using different batches of dye and antibody. These reagents will continually perform with minimal lot-to-lot variation so that you can be confident in your results in a multisite or longitudinal study.

Three overlapping histograms of mouse CD8a BV786
Three overlapping histograms of mouse CD8a BV786

Left: Fluorescence histogram showing mouse CD8a expression identified using three separate lots of BD OptiBuild conjugated antibodies made from different dye lots. Data was derived from gated events with the forward and side light-scatter characteristics of intact splenocytes.

Right: Mouse CD8a expression identified using three separate lots of BD OptiBuild conjugated antibodies made from different dye lots shown against side light-scatter characteristics.

Three overlapping histograms of human CD4 BV421
Three overlapping histograms of human CD4 BV421

Left: Fluorescence histogram showing human CD4 expression identified using three separate lots of BD OptiBuild conjugated antibodies made with different dye lots. Data was derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes.

Right: CD4 expression identified using three separate lots of BD OptiBuild conjugated antibodies made with different dye lots shown vs side light-scatter characteristics.

Three overlapping histograms of mouse CD28 BV650
Three overlapping histograms of mouse CD28 BV650

Left: Fluorescence histogram showing mouse CD28 expression identified using three separate lots of BD OptiBuild conjugated antibodies made with different dye lots. Data was derived from gated events with the forward and side light-scatter characteristics of intact splenocytes.

Right: Mouse CD28 expression identified using three separate lots of BD OptiBuild conjugated antibodies made with different dye lots co-stained with mouse CD4 and mouse CD8a antibodies.

Three overlapping histograms of human CD4 BB700
Three overlapping histograms of human CD4 BB700

Left: Fluorescence histogram showing CD4 expression identified by 3 separate lots of OptiBuild conjugated antibodies. Data was derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes.

Right: CD4 expression identified by 3 separate lots of OptiBuild conjugated antibodies shown against side light-scatter characteristics.

Three overlapping histograms of human CD4 BUV395
Three overlapping histograms of human CD4 BUV395

Left: Fluorescence histogram showing CD4 expression identified by 3 separate lots of OptiBuild conjugated antibodies. Data was derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes.

Right: CD4 expression identified by 3 separate lots of OptiBuild conjugated antibodies shown against side light-scatter characteristics.

Three overlapping histograms of mouse CD8a BV480
Three overlapping histograms of mouse CD8a BV480

Left: Fluorescence histogram showing mouse CD8α expression identified or 3 separate lots of OptiBuild conjugated antibodies was derived from gated events with the forward and side light-scatter characteristics of intact splenocytes.

Right: Mouse CD8α expression identified by 3 separate lots of OptiBuild conjugated antibodies shown against side light-scatter characteristics.

Three overlapping histograms of mouse CD28 BV510
Three overlapping histograms of mouse CD28 BV510

Left: Fluorescence histogram showing mouse CD28 expression identified by 3 separate lots of OptiBuild conjugated antibodies. Data was derived from gated events with the forward and side light-scatter characteristics of intact splenocytes.

Right: Mouse CD28 expression identified by 3 separate lots of OptiBuild conjugated antibodies co-stained with mouse CD4 and mouse CD8a antibodies.

Three overlapping histograms of human CD19 BV605
Three overlapping histograms of human CD19 BV605

Left: Fluorescence histogram showing CD19 expression identified by 3 separate lots of OptiBuild conjugated antibodies. Data was derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes.

Right: CD19 expression identified by 3 separate lots of OptiBuild conjugated antibodies shown versus side light-scatter characteristics.

Three overlapping histograms of mouse CD28 BV650
Three overlapping histograms of mouse CD28 BV650

Left: Fluorescence histogram showing mouse CD28 expression identified by 3 separate lots of OptiBuild conjugated antibodies. Data was derived from gated events with the forward and side light-scatter characteristics of intact splenocytes.

Right: Mouse CD28 expression identified by 3 separate lots of OptiBuild conjugated antibodies co-stained with mouse CD4 and mouse CD8a antibodies.

Three overlapping histograms of human CD4 BV711
Three overlapping histograms of human CD4 BV711

Left: Fluorescence histogram showing CD4 expression identified by 3 separate lots of OptiBuild conjugated antibodies. Data was derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes.

Right:CD4 expression identified by or 3 separate lots of OptiBuild conjugated antibodies shown versus side light-scatter characteristics.

Three overlapping histograms of mouse CD8a BV786
Three overlapping histograms of mouse CD8a BV786

Left: Fluorescence histogram showing mouse CD8a expression identified or 3 separate lots of OptiBuild conjugated antibodies was derived from gated events with the forward and side light-scatter characteristics of intact splenocytes.

Right: Mouse CD8a expression identified by 3 separate lots of OptiBuild conjugated antibodies shown against side light-scatter characteristics.


Leveraging the principles of antigen density and relative fluorochrome brightness, BD OptiBuild reagents have been carefully selected to ensure that they can be readily integrated into your ongoing research. The manufacturing process was rigorously validated to ensure that every conjugation produces an active and specific reagent. As with any custom product, there is a minimal possibility that certain permutations do not perform optimally for biological reasons. If you encounter any issues with the performance of these reagents, please contact our Research Applications team

* BD OptiBuild data on these clone/format combinations is shown for comparative purposes only, and they are not available for sale. There is no overlap between the BD Horizon and BD OptiBuild catalogs.



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