StarBright Dyes

 

 

Image representing StarBright Dye antibody conjugate

Building effective flow cytometry panels requires a wide range of dyes for the excitation lasers of the instrument. With this in mind, we will be offering new StarBright™ Dyes for expanded panel building with our first conjugates being dedicated to the UV laser. Paired together with our extensive antibody clone library, these conjugates will allow researchers to explore new avenues for their next breakthrough.

 

Contact your local representative for assistance in designing and ordering your StarBright panel. For any additional questions, check our FAQ webpage.

 

The image to the right represents a StarBright Dye conjugated to antibodies.
StarBright Dyes are powered by Bio-Rad.

StarBright UltraViolet 575

 

StarBright™ UltraViolet 575 exhibits peak excitation/emission wavelengths at 340 nm and 569 nm respectively, most closely matching that of BD Horizon™ BUV563. On conventional cytometers, it can be detected with a 355 nm ultraviolet laser and a 586/15 filter set (or equivalent). With proper panel building, it can be used together in panels with StarBright UV740, StarBright UV795, and Spark PLUS UV395™. With its high brightness, StarBright UV575 is best suited for detecting antigens with low to moderate expression levels. In addition, StarBright UV575 can be cocktailed with other fluorophore conjugated antibodies without exhibiting inter-dye interactions. It is also stable towards exposure to heat or a variety of commonly used fixative solutions.

 

 

Excitation and Emission Spectra of StarBright UltraViolet 575

 

StarBright UV575 spectral signature

 

Emission spectra (top) and normalized emission spectra (middle) of StarBright UV575 run on a 5-laser Cytek™ Aurora Spectral Cytometer. To compare StarBright UV575 with other fluorophores on a spectral cytometer, use our Aurora Spectral Analyzer tool.

 

Normalized excitation and emission spectra (bottom) of StarBright UV575 obtained from a spectrophotometer. To compare StarBright UV575 with other fluorophores, use our Fluorescence Spectra Analyzer tool.

 

Go Beyond BD Horizon™ BUV563

 

StarBright UV575 comparison to BD BUV563

 

Human peripheral blood lymphocytes were stained with anti-human CD3 (clone UCHT1) Spark Violet™ 423 and anti-human CD4 (clone SK3) conjugated to StarBright UV575 and compared BD Horizon™ BUV563. Samples were acquired on a 5-laser Cytek® Aurora.

 

Multicolor Staining With StarBright UltraViolet 575

 

Panel A demonstrates how StarBright UV575 performs in a multicolor panel with fluorophores that share significant spectra overlap with it. Panel B is shown as a reference panel using pre-optimized fluorophores to ensure that staining patterns are similar.

 

StarBright UV575 Panel

 

 

Marker

Panel A Fluorophore

Panel B Fluorophore

CD4

StarBright UV575

StarBright UV575

CD8

StarBright UV795

Pacific Blue

CD19

Spark PLUS UV395™

FITC

CD56

StarBright UV740

PE

 

Human PBMCs were stained with the indicated antibodies and analyzed on a spectral flow cytometer. All plots are gated on lymphocytes.

 

 

Stability and Validation Testing

 

All BioLegend fluorophores undergo rigorous testing procedures to determine how light, heat, and fixation may affect the performance and ensure they will perform reliably. To compare the signal across different conditions and timepoints, we used the Stain Index (formula below) to measure the relative brightness of the antibody. 

Spark 550

Photostability Testing

 

The photostability of StarBright UV575 was tested in two ways that mimic how an antibody may be exposed to light over the course of an experiment.

 

  1. Antibodies were stored in the dark or exposed to fluorescent lighting. Then, the antibodies were used to stain freshly harvested cell samples and analyzed immediately.
  2. Cells were stained with antibody that had been kept under recommended storage conditions. Prior to analysis, the stained cells were stored in the dark or exposed to fluorescent lighting.

 

StarBright UV575 Photostability

 

To assess the photostability of StarBright UV575-conjugated antibodies, anti-human CD4 (clone SK3) StarBright UV575 was exposed to light or protected in the dark for the indicated times (AB Only). The samples were then used to stain human PBMCs. To measure the photostability of StarBright UV575 stained cells, human PBMCs were stained with anti-human CD4 (clone SK3) StarBright UV575 (+ Cells). Stained cells were exposed to light or kept in the dark as indicated and the staining index of lymphocytes was calculated.

 

Heat Stability

 

StarBright UV575 Heat Stability

Anti-human CD4 (clone SK3) StarBright UV575 was aliquoted and incubated at the indicated temperatures over the course of 28 days. The antibodies were then used to stain human lysed whole blood from a single donor and the staining index was calculated.

 

Fixative Stability

 

A guide to the fixatives used in this experiment:

 

 

StarBright UV575 Fixation Data

 

Human PBMCs were stained with anti-human CD4 (clone SK3) conjugated to StarBright UV575 and fixed using the respective protocols for each buffer set. Fresh samples were fixed and read on a 5-laser Cytek™ Aurora spectral cytometer immediately. Overnight samples were fixed and stored in Cell Staining Buffer overnight before acquisition. Data shown was gated on lymphocytes.

 

StarBright UltraViolet 575

 

StarBright™ UltraViolet 575 exhibits peak excitation/emission wavelengths at 340 nm and 569 nm respectively, most closely matching that of BD Horizon™ BUV563. On conventional cytometers, it can be detected with a 355 nm ultraviolet laser and a 586/15 filter set (or equivalent). With proper panel building, it can be used together in panels with StarBright UV740, StarBright UV795, and Spark PLUS UV395™. With its high brightness, StarBright UV575 is best suited for detecting antigens with low to moderate expression levels. In addition, StarBright UV575 can be cocktailed with other fluorophore conjugated antibodies without exhibiting inter-dye interactions. It is also stable towards exposure to heat or a variety of commonly used fixative solutions.

 

 

Excitation and Emission Spectra of StarBright UltraViolet 575

 

StarBright UV575 spectral signature

 

Emission spectra (top) and normalized emission spectra (middle) of StarBright UV575 run on a 5-laser Cytek™ Aurora Spectral Cytometer. To compare StarBright UV575 with other fluorophores on a spectral cytometer, use our Aurora Spectral Analyzer tool.

 

Normalized excitation and emission spectra (bottom) of StarBright UV575 obtained from a spectrophotometer. To compare StarBright UV575 with other fluorophores, use our Fluorescence Spectra Analyzer tool.

 

Go Beyond BD Horizon™ BUV563

 

StarBright UV575 comparison to BD BUV563

 

Human peripheral blood lymphocytes were stained with anti-human CD3 (clone UCHT1) Spark Violet™ 423 and anti-human CD4 (clone SK3) conjugated to StarBright UV575 and compared BD Horizon™ BUV563. Samples were acquired on a 5-laser Cytek® Aurora.

 

Multicolor Staining With StarBright UltraViolet 575

 

Panel A demonstrates how StarBright UV575 performs in a multicolor panel with fluorophores that share significant spectra overlap with it. Panel B is shown as a reference panel using pre-optimized fluorophores to ensure that staining patterns are similar.

 

StarBright UV575 Panel

 

 

Marker

Panel A Fluorophore

Panel B Fluorophore

CD4

StarBright UV575

StarBright UV575

CD8

StarBright UV795

Pacific Blue

CD19

Spark PLUS UV395™

FITC

CD56

StarBright UV740

PE

 

Human PBMCs were stained with the indicated antibodies and analyzed on a spectral flow cytometer. All plots are gated on lymphocytes.

 

 

Stability and Validation Testing

 

All BioLegend fluorophores undergo rigorous testing procedures to determine how light, heat, and fixation may affect the performance and ensure they will perform reliably. To compare the signal across different conditions and timepoints, we used the Stain Index (formula below) to measure the relative brightness of the antibody. 

Spark 550

Photostability Testing

 

The photostability of StarBright UV575 was tested in two ways that mimic how an antibody may be exposed to light over the course of an experiment.

 

  1. Antibodies were stored in the dark or exposed to fluorescent lighting. Then, the antibodies were used to stain freshly harvested cell samples and analyzed immediately.
  2. Cells were stained with antibody that had been kept under recommended storage conditions. Prior to analysis, the stained cells were stored in the dark or exposed to fluorescent lighting.

 

StarBright UV575 Photostability

 

To assess the photostability of StarBright UV575-conjugated antibodies, anti-human CD4 (clone SK3) StarBright UV575 was exposed to light or protected in the dark for the indicated times (AB Only). The samples were then used to stain human PBMCs. To measure the photostability of StarBright UV575 stained cells, human PBMCs were stained with anti-human CD4 (clone SK3) StarBright UV575 (+ Cells). Stained cells were exposed to light or kept in the dark as indicated and the staining index of lymphocytes was calculated.

 

Heat Stability

 

StarBright UV575 Heat Stability

Anti-human CD4 (clone SK3) StarBright UV575 was aliquoted and incubated at the indicated temperatures over the course of 28 days. The antibodies were then used to stain human lysed whole blood from a single donor and the staining index was calculated.

 

Fixative Stability

 

A guide to the fixatives used in this experiment:

 

 

StarBright UV575 Fixation Data

 

Human PBMCs were stained with anti-human CD4 (clone SK3) conjugated to StarBright UV575 and fixed using the respective protocols for each buffer set. Fresh samples were fixed and read on a 5-laser Cytek™ Aurora spectral cytometer immediately. Overnight samples were fixed and stored in Cell Staining Buffer overnight before acquisition. Data shown was gated on lymphocytes.

 

StarBright UltraViolet 740

 

StarBright™ UltraViolet 740 exhibits peak excitation/emission wavelengths at 343 nm and 737 nm respectively, most closely matching that of BD Horizon™ BUV737. On conventional cytometers, it can be detected with a 355 nm ultraviolet laser and a 740/35 filter set (or equivalent). With proper panel building, it can be used together in panels with StarBright UV575, StarBright UV795, and Spark PLUS UV395™. This extremely bright fluorophore is best suited for detecting antigens with low expression levels. In addition, StarBright UV740 can be cocktailed with other fluorophore conjugated antibodies without exhibiting inter-dye interactions. It is also stable towards exposure to heat or a variety of commonly used fixative solutions.

 

 

Excitation and Emission Spectra of StarBright UltraViolet 740

 

StarBright UV740 spectral signature

 

Emission spectra (top) and normalized emission spectra (middle) of StarBright UV740 run on a 5-laser Cytek™ Aurora Spectral Cytometer. To compare StarBright UV740 with other fluorophores on a spectral cytometer, use our Aurora Spectral Analyzer tool.

 

Normalized excitation and emission spectra (bottom) of StarBright UV740 obtained from a spectrophotometer. To compare StarBright UV740 with other fluorophores, use our Fluorescence Spectra Analyzer tool.

 

Go Beyond BD Horizon™ BUV737

 

StarBright UV740 Comparison Against BD BUV737

 

Human peripheral blood lymphocytes were stained with anti-human CD3 (clone UCHT1) Spark Violet™ 423 and anti-human CD4 (clone SK3) conjugated to StarBright UV740 and compared against BD Horizon™ BUV737. Samples were acquired on a 5-laser Cytek® Aurora.

 

Multicolor Staining With StarBright UltraViolet 740

 

Panel A demonstrates how StarBright UV740 performs in a multicolor panel with fluorophores that share significant spectra overlap with it. Panel B is shown as a reference panel using pre-optimized fluorophores to ensure that staining patterns are similar.

 

StarBright UV740 Panel

 

 

Marker

Panel A Fluorophore

Panel B Fluorophore

CD4

StarBright UV740

StarBright UV740

CD8

StarBright UV575

Pacific Blue

CD19

StarBright UV795

FITC

CD56

Spark PLUS UV395™

PE

 

Human peripheral blood cells were stained with the indicated antibodies and analyzed on a spectral flow cytometer. All plots are gated on lymphocytes.

 

 

Stability and Validation Testing

 

All BioLegend fluorophores undergo rigorous testing procedures to determine how light, heat, and fixation may affect the performance and ensure they will perform reliably. To compare the signal across different conditions and timepoints, we used the Stain Index (formula below) to measure the relative brightness of the antibody. 

Spark 550

Photostability Testing

 

The photostability of StarBright UV740 was tested in two ways that mimic how an antibody may be exposed to light over the course of an experiment.

 

  1. Antibodies were stored in the dark or exposed to fluorescent lighting. Then, the antibodies were used to stain freshly harvested cell samples and analyzed immediately.
  2. Cells were stained with antibody that had been kept under recommended storage conditions. Prior to analysis, the stained cells were stored in the dark or exposed to fluorescent lighting.

 

StarBright UV740 Photostability

 

To assess the photostability of StarBright UV740-conjugated antibodies, anti-human CD4 (clone SK3) StarBright UV740 was exposed to light or protected in the dark for the indicated times (AB Only). The samples were then used to stain human PBMCs. To measure the photostability of StarBright UV740 stained cells, human PBMCs were stained with anti-human CD4 (clone SK3) StarBright UV740 (+ Cells). Stained cells were exposed to light or kept in the dark as indicated and the staining index of lymphocytes was calculated.

 

Heat Stability

 

StarBright UV740 Heat Stability

Anti-human CD4 (clone SK3) StarBright UV740 was aliquoted and incubated at the indicated temperatures over the course of 28 days. The antibodies were then used to stain human lysed whole blood from a single donor, and the staining index of lymphocytes was calculated.

 

Fixative Stability

 

A guide to the fixatives used in this experiment:

 

 

StarBright UV740 Fixation Data

 

Human PBMCs were stained with anti-human CD4 (clone SK3) conjugated to StarBright UV740 and fixed using the respective protocols for each buffer set. Fresh samples were fixed and read on a 5-laser Cytek™ Aurora spectral cytometer immediately. Overnight samples were fixed and stored in Cell Staining Buffer overnight before acquisition. Data shown was gated on lymphocytes.

 

StarBright UltraViolet 795

 

StarBright™ UltraViolet 795 exhibits peak excitation/emission wavelengths at 340 nm and 792 nm respectively, most closely matching that of BD Horizon™ BUV805. On conventional cytometers, it can be detected with a 355 nm ultraviolet laser and a 820/60 filter set (or equivalent). With proper panel building, it can be used together in panels with StarBright UV575, StarBright UV740, and Spark PLUS UV395™. This extremely bright fluorophore is best suited for detecting antigens with low expression levels. In addition, StarBright UV795 can be cocktailed with other fluorophore conjugated antibodies without exhibiting inter-dye interactions. It is also stable towards exposure to heat or a variety of commonly used fixative solutions.

 

 

Excitation and Emission Spectra of StarBright UltraViolet 795

 

StarBright UV795 spectral signature

 

Emission spectra (top) and normalized emission spectra (middle) of StarBright UV795 run on a 5-laser Cytek™ Aurora Spectral Cytometer. To compare StarBright UV795 with other fluorophores on a spectral cytometer, use our Aurora Spectral Analyzer tool.

 

Normalized excitation and emission spectra (bottom) of StarBright UV795 obtained from a spectrophotometer. To compare StarBright UV795 with other fluorophores, use our Fluorescence Spectra Analyzer tool.

 

Go Beyond BD Horizon™ BUV805

 

StarBright UV795 comparison data against BD BUV805

 

Human peripheral blood lymphocytes were stained with anti-human CD3 (clone UCHT1) Spark Violet™ 423 and anti-human CD4 (clone SK3) conjugated to StarBright UV795 and compared against BD Horizon™ BUV805. Samples were acquired on a 5-laser Cytek® Aurora.

 

Multicolor Staining With StarBright UltraViolet 795

 

Panel A demonstrates how StarBright UV795 performs in a multicolor panel with fluorophores that may exhibit spectral overlap with it. Panel B is shown as a reference panel using pre-optimized fluorophores to ensure that staining patterns are similar.

 

StarBright UV759 Panel

 

 

Marker

Panel A Fluorophore

Panel B Fluorophore

CD4

StarBright UV795

StarBright UV795

CD8

StarBright UV575

Pacific Blue

CD19

Spark PLUS UV395™

FITC

CD56

StarBright UV740

PE

 

Human peripheral blood cells were stained with the indicated antibodies and analyzed on a spectral flow cytometer. All plots are gated on lymphocytes.

 

Stability and Validation Testing

 

All BioLegend fluorophores undergo rigorous testing procedures to determine how light, heat, and fixation may affect the performance and ensure they will perform reliably. To compare the signal across different conditions and timepoints, we used the Stain Index (formula below) to measure the relative brightness of the antibody. 

Photostability Testing

 

The photostability of StarBright UV795 was tested in two ways that mimic how an antibody may be exposed to light over the course of an experiment.

 

  1. Antibodies were stored in the dark or exposed to fluorescent lighting. Then, the antibodies were used to stain freshly harvested cell samples and analyzed immediately.
  2. Cells were stained with antibody that had been kept under recommended storage conditions. Prior to analysis, the stained cells were stored in the dark or exposed to fluorescent lighting.

 

StarBright UV795 Photostability

 

To assess the photostability of StarBright UV795-conjugated antibodies, anti-human CD4 (clone SK3) StarBright UV795 conjugates were exposed to light or protected in the dark (Ab Only). The samples were then used to stain human PBMCs. To measure the photostability of StarBright UV795 stained cells, human PBMCs were stained with anti-human CD4 (clone SK3) StarBright UV795 (+ Cells). Stained cells were exposted to light or kept in the dark as indicated, and the staining index of lymphocytes was calculated. 

 

Heat Stability

 

Anti-human CD4 (clone SK3) StarBright UV795 was aliquoted and incubated at the indicated temperatures over the course of 28 days. The antibodies were then used to stain human lysed whole blood from a single donor, and the staining index of lymphocytes was calculated.

 

Fixative Stability

 

A guide to the fixatives used in this experiment:

 

 

StarBright UV795 Fixation Data

 

Human PBMCs were stained with anti-human CD4 (clone SK3) conjugated to StarBright UV795 and fixed using the respective protocols for each buffer set. Fresh samples were fixed and read on a 5-laser Cytek™ Aurora spectral cytometer immediately. Overnight samples were fixed and stored in Cell Staining Buffer overnight before acquisition. Data shown was gated on lymphocytes.

 

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