Flow cytometry is a core application for most immunologists, so using the best reagents produces the best results. BioLegend has proven our commitment to bringing the highest quality and support to flow cytometry reagents at the best value on the market. We provide an outstanding selection of flow cytometry buffers and fluorophore options for our antibodies, including high performance Brilliant Violet™ conjugates and our featured formats: PE/Dazzle™ 594 and APC/Fire™ 750.

Immunology – Flow Cytometry Primary Antibodies

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Learn more about our fluorophore options


Available fluorophore-antibody conjugates by laser.

Click to view spectral data.

      Violet (405 nm)

  • Brilliant Violet 421™
  • Pacific Blue™
  • Brilliant Violet 510™
  • Brilliant Violet 570™
  • Brilliant Violet 605™
  • Brilliant Violet 650™
  • Brilliant Violet 711™
  • Brilliant Violet 785™

      Blue (488 nm)

  • FITC
  • Alexa Fluor® 488
  • PE
  • PE/Dazzle™ 594
  • PE/Cy5
  • PerCP
  • PerCP/Cy5.5
  • PE/Cy7
  • Spark Blue™ 550

      Yellow/Green
     (532-561 nm)

  • PE
  • PE/Dazzle™ 594
  • PE/Cy5
  • PE/Cy7

      Red (635 nm)

  • APC
  • Alexa Fluor® 647
  • Alexa Fluor® 700
  • APC/Cy7
  • APC/Fire™ 750
  • Spark NIR™ 685

Learn more about multicolor flow cytometry


Webinars and Videos

 

 

 

 

Tutorial Pages

Immunobiology Handbook Chapters

 

  • Build a Better Multicolor Flow Cytometry Assay
  • Buffers for Flow Cytometry
  • Cell Health, Cell Cycle and Proliferation
  • Antibodies for Non-Human Primate (NHP) Research
  • Cell Separation
  • MHC Multimers
  • Neuroinflammation
  • Protocols

Violet (405 nm)

 

Fluorophore Brightness Index (1=dim, 5=bright) Exmax Emmax Comments
Brilliant Violet 421™ 4 405 nm 421 nm Brilliant Violet 421™ is a novel organic fluorescent polymer capable of extremely bright signal and high photostability. It is an excellent choice for flow cytometry and microscopy. It is not recommended for use with Pacific Blue™.
Pacific Blue™ 1 401 nm 455 nm Pacific Blue™ is a hydroxycoumarin-based fluorophore, generally considered to be dimmer than most other fluorophores. It is recommended for highly expressed antigens. Brilliant Violet 421™ is a brighter alternative for weakly expressed antigens. 
Brilliant Violet 510™ 1 405 nm 510 nm Brilliant Violet 510™ is a novel polymer with excellent signal-to-noise, excited by the violet laser. It can provide dramatic improvements over existing fluorophores emitting in this range such as Pacific Orange™ and Horizon™ V500.
Brilliant Violet 570™ 1 405 nm 570 nm Brilliant Violet 570™ is a novel organic fluorescent polymer. It provides a much brighter alternative to Pacific Orange™ for multicolor flow on the violet laser and is a better alternative to nanocrystals for intracellular flow cytometry.
Brilliant Violet 605™ 3 405 nm 603 nm Brilliant Violet 605™ is a novel organic fluorescent polymer. It provides a much brighter alternative to eFluor® 605 for multicolor flow on the violet laser and is a better alternative to nanocrystals for intracellular flow cytometry.
Brilliant Violet 650™ 3 405 nm 646 nm Brilliant Violet 650™ is a novel organic fluorescent polymer. It provides a much brighter alternative to eFluor® 650 for multicolor flow on the violet laser and is a better alternative to nanocrystals for intracellular flow cytometry.
Brilliant Violet 711™ 4 405 nm 711 nm Brilliant Violet 711™ is a novel molecule based on the Brilliant Violet 421™ polymer core. It provides a much brighter alternative to eFluor® 700NC for multicolor flow on the violet laser and is a better alternative to nanocrystals for intracellular flow cytometry.
Brilliant Violet 750™ 3 405 nm 750 nm Brilliant Violet 750™ is a novel molecule based on the Brilliant Violet 421™ polymer core. It provides further options for the violet laser, particularly for those with either a spectral detection cytometer or a cytometer with a decagon configuration for the violet laser. Alternatively, it can be used in place of BV785™ on a standard violet laser octagon configuration.
Brilliant Violet 785™ 3 405 nm 785 nm Brilliant Violet 785™ is a novel molecule based on the Brilliant Violet 421™ polymer core. It provides further options for the violet laser and is a better alternative to nanocrystals for intracellular flow cytometry.

Blue Laser (488 nm)

 

Fluorophore Brightness Index (1=dim, 5=bright) Exmax Emmax Comments
Alexa Fluor® 488 1 495 nm 519 nm Alexa Fluor® 488 is a fluorescent molecule with exceptional photostability. For flow cytometry it has very similar brightness and emission properties compared to FITC, but is advantageous for microscopy applications. 
FITC 1 494 nm 520 nm FITC (fluorescein isothiocyanate) is a small synthetic organic fluorophore with moderate brightness, suitable for flow and microscopy applications. It is sensitive to pH changes. It cannot be used with Alexa Fluor® 488.
PerCP 1 482 nm 678 nm PerCP is a naturally-derived carotenoid pigment found in photosynthetic dinoflagellates. It is not recommended for high powered lasers >25 mW due to its sensitivity to photobleaching. PerCP has overlapping emission with PE/Cy5, so these should not be used on the same laser.
PerCP/Cy5.5
PerCP/Cyanine5.5
2 482 nm 695 nm PerCP/Cyanine5.5 is tandem conjugate of PerCP and the cyanine dye, Cyanine5.5. Unlike PerCP, it is quite photostable and can be used with high powered lasers. Of the all tandems, it is the most photostable and resistant to degradation by fixation. PerCP and PerCP/Cyanine5.5 have significant overlap and are not recommended for use together.
Spark Blue™ 550 1 516 nm 550 nm Spark Blue™ 550 is designed to be used in high level (>22 color) flow cytometry assays on a spectral unmixing cytometer. It is excited off of the 488 nm blue laser and emits between the peak emission of FITC and PE. Emission off the 405 nm violet laser adds to the accuracy with which the fluorophore can be unmixed. It can be used as an alternative to Alexa Fluor® 532.
PE 5 496 nm 578 nm R-Phycoerythrin (PE) is a pigment derived from red algae. It is a large 240 kD protein that contains 23 phycoerthrobilin chromophores per molecule. It is extremely bright for flow cytometry, but poor photostability makes it unsuitable for microscopy. PE and its tandem are also sensitive to denaturation by fixation. Because of its wide excitation range, it can be excited by the 488, 532, and 561 nm lasers.
PE/Dazzle™ 594 5 565 nm 610 nm PE/Dazzle™ 594 is a tandem conjugate of PE and BioLegend’s proprietary fluorophore DZL594. Similar to PE, it is extremely bright for flow cytometry. It is spectrally similar to PE-Texas Red®, BD Horizon™ PE-CF594, and PE-eFluor® 610.
PE/Cy5 5 496 nm 667 nm PE/Cy5 is a tandem conjugate of PE and the cyanine dye, Cy5. Similar to PE, it is bright for flow cytometry, but is sensitive to photobleaching and not recommended for microscopy. Cy5 has been known to non-specifically bind to Fc receptors. On some instruments, it is not recommended for use with APC, due to their overlapping emissions.
PE/Cy7 4 496 nm 785 nm PE/Cy7 is a tandem conjugate of PE and the cyanine dye, Cy7. Similar to PE, it is bright for flow cytometry, but is sensitive to photobleaching and not recommended for microscopy. This tandem is particularly sensitive to light exposure and prolonged fixation.

Green laser (532 nm)/Yellow-Green laser (561 nm)

 

Fluorophore Brightness Index (1=dim, 5=bright) Exmax Emmax Comments
PE 5 496 nm 578 nm R-Phycoerythrin (PE) is a pigment derived from red algae. It is a large 240 kD protein that contains 23 phycoerthrobilin chromophores per molecule. It is extremely bright for flow cytometry, but poor photostability makes it unsuitable for microscopy. PE and its tandem are also sensitive to denaturation by fixation. Because of its wide excitation range, it can be excited by the 488, 532, and 561 nm lasers.
PE/Dazzle™ 594 5 565 nm 610 nm PE/Dazzle™ 594 is a tandem conjugate of PE and BioLegend’s proprietary fluorophore DZL594. Similar to PE, it is extremely bright for flow cytometry. It is spectrally similar to PE-Texas Red®, BD Horizon™ PE-CF594, and PE-eFluor® 610.
PE/Cy5 5 496 nm 667 nm PE/Cy5 is a tandem conjugate of PE and the cyanine dye, Cy5. Similar to PE, it is bright for flow cytometry, but is sensitive to photobleaching and not recommended for microscopy. Cy5 has been known to non-specifically bind to Fc receptors. On some instruments, it is not recommended for use with APC, due to their overlapping emissions.
PE/Cy7 4 496 nm 785 nm PE/Cy7 is a tandem conjugate of PE and the cyanine dye, Cy7. Similar to PE, it is bright for flow cytometry, but is sensitive to photobleaching and not recommended for microscopy. This tandem is particularly sensitive to light exposure and prolonged fixation.

Red laser (633 nm)

 

Fluorophore Brightness Index (1=dim, 5=bright) Exmax Emmax Comments
APC 4 650 nm 660 nm Allophycocyanin (APC) is a 105 kD molecule derived from blue-green algae that has 6 phycocyanobilin chromophores per molecule. It is bright and particularly useful for flow cytometry but not microscopy. It should not be used together with Alexa Fluor® 647 due to their overlapping emissions.
Alexa Fluor® 647 4 650 nm 668 nm Alexa Fluor® 647 is spectrally equivalent to APC, but has much better photostability, making it ideal for microscopy applications. While generally not as bright as APC, it is also a good choice for flow cytometry.
Spark NIR™ 685 2 660 nm 685 nm Spark NIR™ 685 is designed to be used in high level (>22 color) flow cytometry assays on a spectral unmixing cytometer. It emits between the peak emission of Alexa Fluor® 647 and Alexa Fluor® 700. It can be used in the same panel as Alexa Fluor® 647 but ideally, they should not be used to detect two co-expressed markers.
Alexa Fluor® 700 2 696 nm 719 nm Althought it is a dim molecule, Alexa Fluor® 700, expands the choices for the red laser, fitting nicely between APC and APC/Cy7.
APC/Cy7 2 650 nm 785 nm APC/Cy7 is a tandem of APC and the cyanin dye, Cy7. It is particularly sensitive to light and fixation and should be handled with care to avoid light exposure.
APC/Fire™ 750 2 650 nm 787 nm APC/Fire™ 750 is a new replacement for APC/Cy7 with improved temperature and photostability. APC/Fire™ 750 also has lower compensation requirements than APC/Cy7 conjugates while maintaining an equal level of brightness. Additionally, APC/Fire™ 750 has minimal non-specific binding to monocytes, as has been observed with APC/Cy7. Lastly, APC/Fire™ 750 conjugates are consistently brighter than APC-H7 in all conjugates tested.