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

More Flow Cytometry Reagents

Browse the Immunobiology Catalog by Other Applications

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

 

 

 

 

 

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.

BioLegend now brings exceptional value and performance for cell separation with MojoSort™, our magnetic cell separation system.

MojoSort™ nanobeads are BioLegend’s proprietary magnetic particles conjugated to antibodies or streptavidin. They are suitable for use in our MojoSort™ magnet, a tube-based magnetic device, as well as other commercial tube-based magnetic devices and magnetic columns.

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MojoSort Videos

Antibodies can be used for a variety of functional assays, including blocking, activation, depletion, or neutralization, for both in vitro and in vivo purposes. It is critical that these reagents are free of preservatives, such as sodium azide, and have very low levels of endotoxin. BioLegend offers biofunctional antibodies in three product formats: LEAF™ (Low Endotoxin, Azide-Free), Ultra-LEAF™, and GoInvivo™. Compare the formats below.

intro image
  GoInVivo™ Ultra-LEAF™ LEAF™
Endotoxin Level < 1 EU/mg of protein < 10 EU/mg of protein < 100 EU/mg of protein
Sizing Options 5 mg - 1 g 100 µg - 25 mg 50 µg- 1 mg
QC Testing Flow Cytometry Flow Cytometry Flow Cytometry
Pricing Best value per mg Better value per mg Good value
Pathogen Testing Yes No No
Functional Validation Yes No No
Number of Products 20+ 100+ 320+
Notes Focused on immune checkpoint receptor antibodies Excellent for in vivo administration  

 

Featured Products


 

GoInVivo™ Purified anti-mouse CD274 (B7-H1, PD-L1)

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CD274, also known as B7-H1 or programmed death ligand 1 (PD-L1), is a 40 kD type I transmembrane protein and a member of the B7 family within the immunoglobulin receptor superfamily. It is expressed on T cells, B cells, NK cells, dendritic cells, IFN-γ activated endothelial cells, and monocytes. B7-H1 is one of the ligands of PD-1. The interaction of B7-H1 with PD-1 plays an important role in the inhibition of T cell responses. Other studies have shown that B7-H1 is able to costimulate T cell growth and cytokine production. CD274 is involved in costimulation essential for T cell proliferation and production of IL-10 and IFN-γ, in an IL-2-dependent and a PD-1-independent manner. Its interaction with PD-1 inhibits T cell proliferation and cytokine production.

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Treg-Protector™ (anti-ARTC2 Nanobody)

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ARTC2, also known as ART2.2, is a 35 kD, GPI-anchored, and cell surface ADP-ribosyltransferase that is highly expressed by regulatory T cells (Tregs) and natural killer T (NKT) cells. ARTC2 is enzymatically active at 4°C. Through the ribosylation of the ion channel P2X7, ARTC2 mediates NAD+-induced cell death (NICD) and the proteolytic shedding of CD27 and CD62L. Blocking the activity ARTC2 with Treg-Protector™ improves the recovery of Tregs and NKT cells during their isolation from lymphatic and peripheral tissues.

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LEAF™ Purified anti-mouse DR3 (TNFRSF25)

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DR3, also known as TNFRSF25, lymphocyte-associated receptor of death (LARD), WSL-1, TRAMP, TR3 and APO3, is a 55 kD glycoprotein belonging to the TNF receptor superfamily (TNFRSF). TNFSF25 is expressed at low level on CD4-positive, CD8-positive T cells and NKT cells. It has been shown to stimulate NF-κB activity and regulate cell apoptosis. The signal transduction of this receptor is mediated by various death domains containing adaptor proteins. Recently, it has been shown that TNFRSF25 is also expressed on Treg cells and regulates Treg proliferation.

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Ultra-LEAF™ Purified anti-mouse CX3CR1

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CX3CR1 is a 40 kD, G-protein coupled receptor, with seven transmembrane regions. CX3CR1 is expressed by resident and alternatively activated macrophages (M2), a subset of monocytes, dendritic cells (DCs), NK cells, a subset of memory T cells, and mast cells. CX3CR1 is involved in cell recruitment during inflammation and participates in cell adhesion and extravasation from blood vessels. Its ligand is CX3CL1, also known as fractalkine or neurotactin. CX3CR1 is also a coreceptor for HIV1 and variations in this gene leads to increased susceptibility to HIV. In the brain, it is expressed by glial cells, which interact with CX3CL1 expressed by neurons.

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Ultra-LEAF™ Purified anti-mouse CD182 (CXCR2)

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CD182, also known as CXCR2, is a G-protein coupled receptor with seven transmembrane regions that is involved in chemotaxis, neutrophil activation, and angiogenesis. CXCR2 is expressed by neutrophils, basophils, a subset of T cells, monocytes, macrophages, natural killer (NK) and natural killer T cells (NKT). Its ligands are CXCL1, CXCL2, CXCL3, and CXCL5.

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BioLegend's LEGENDScreen™ products are lyophilized, fluorophore-conjugated antibodies provided in 96-well plates for the purpose of screening cell surface molecules on your cells of interest.

These antibody plates provide a large selection of specificities per screen: The Human Cell Screening Kit has 332 specificities plus 10 isotype controls, while the Mouse Cell Screening Kit has 252 cell surface marker antibodies plus 11 isotype controls. Directly conjugated antibodies at pre-titrated optimal concentrations provide reliable results. Each kit also comes with staining buffer, fixation buffer, and plate sealers.

Lscreen Intro Image

Citations

 

 

Product Authors Journal Year Vol/Page Pubmed
LEGENDScreen™ Human Cell Screening (PE) Kit Schütz F, Hackstein H Biochem Biophys Res Commun 2014 443:453 Link
LEGENDScreen™ Human Cell Screening (PE) Kit Graessel A, et al. Mol Cell Proteomics 2015 14: 2085-2102 Link
LEGENDScreen™ Human Cell Screening (PE) Kit Koh P, et al. Sci Data 2016 3:160109 Link
LEGENDScreen™ Human Cell Screening (PE) Kit Collier A, et al. Cell Stem Cell 2017 10.1016/j.stem.2017.02.014 Link
LEGENDScreen™ Human PE Kit Bojic S, et al. Stem Cells 2018 36:1723 Link
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TotalSeq™ – BioLegend’s oligonucleotide conjugated antibodies

 

While traditional flow cytometry allows for rapid, multi-dimensional characterization of cell populations, it lacks the resolution and capacity to perform simultaneous proteomic and genomic analysis on a single-cell scale. BioLegend now offers oligonucleotide-conjugated TotalSeq™ antibodies designed for single-cell proteogenomic analysis via cellular indexing of transcriptomes and epitopes by sequencing, or CITE-Seq. They are also compatible with similar workflows, such as REAP-seq.


In place of fluorophores, each TotalSeq™ clone is conjugated to an oligonucleotide that includes a unique 15 nucleotide (nt) barcode, which converts antigen signal into a readable sequence. Combining antigen detection with single cell RNA sequencing (scRNA-Seq) allows for simultaneous transcriptomic and proteomic analyses in individual cells. Furthermore, since fluorophore and detector availabilities are no longer a factor, a significantly larger number of markers that can be analyzed at once. So far, simultaneous analysis of over 80 surface markers with scRNA-Seq has been reported in the literature.


In addition to unique antigen-specific antibodies, we also carry oligo-conjugated cell hashing reagents, which contain antibodies to ubiquitously expressed cell markers to facilitate sample multiplexing and doublet exclusion. TotalSeq™ antibodies are readily compatible with 10X Genomics scRNA-Seq and Illumina sequencing platforms, but are also adaptable to other droplet-based single-cell workflows with assay optimization.

General TotalSeq™ antibody schematic:

 

The oligonucleotides conjugated to our TotalSeq™ antibodies have 3 main components: a 5’ PCR handle, followed by a 15 nt barcode unique to the clone, and a 3’ flanking sequence. The 3’ sequence determines compatibility to different scRNA-Seq assays. The example shown above is a schematic for our TotalSeq™-A antibody, which contains a Poly-A stretch on the 3’ end. They are compatible with 10X Genomics 3’ kit (v2), but also with any other system that employs poly-A mRNA capture.

Additional resources:

  • CITE-seq | A collection of resources for performing CITE-seq
  • Simultaneous epitope and transcriptome measurement in single cells: Stoeckius M, et al. 2017. Nat. Methods. 14(9):865-868 PubMed
  • Cell "hashing" with barcoded antibodies enables multiplexing and doublet detection for single cell genomics: Stoeckius M, et al. 2017 bioRxiV Link
  • Multiplexed quantification of proteins and transcripts in single cells: Peterson VM, et al. 2017. Nat. Biotechnol. 35(10):936-939 PubMed
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