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Brilliant violet 650

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Brilliant Violet 650 is a fluorescent dye used in flow cytometry applications. It has an excitation maximum at 405 nm and an emission maximum at 650 nm. The dye can be used to label antibodies or other biomolecules for detection and analysis in flow cytometry experiments.

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Lab products found in correlation

Brilliant violet 421, by BioLegend (7 mentions) Brilliant violet 605, by BioLegend (3 mentions) Alexa fluor 647, by BioLegend (3 mentions) Permeabilization buffer, by Thermo Fisher Scientific (2 mentions) Ber act8, by BioLegend (2 mentions) Fixation buffer, by Thermo Fisher Scientific (2 mentions) Pe texas red, by Thermo Fisher Scientific (2 mentions) Sj25 c1, by Thermo Fisher Scientific (2 mentions) Percpfl710, by Thermo Fisher Scientific (2 mentions) Uchl1, by Thermo Fisher Scientific (2 mentions) 6 laser lsrii analytical flow cytometer, by BD (2 mentions) Alexa fluor 488, by BioLegend (2 mentions) Fluorescein isothiocyanate (fitc), by BD (2 mentions) Apc cy7, by BD (2 mentions) Brilliant violet 510, by BioLegend (2 mentions) Alexa fluor 647, by Thermo Fisher Scientific (1 mentions) Facsaria 2, by BD (1 mentions) Pe anti mouse cd11c, by BioLegend (1 mentions) Biotinylated peanut agglutinin pna, by Vector Laboratories (1 mentions) Alexa fluor 488 anti mouse rat human foxp3 antibody, by BioLegend (1 mentions)

Close spelling variants of this product

Anti-CD4 Brilliant Violet 650 (3 citations) Brilliant Violet 650 anti-human EPCAM (2 citations) CD4-Brilliant Violet 650 (clone OKT4; (3 citations) CD19-Brilliant Violet 650 (2 citations) Brilliant violet 650-conjugated CD4 (3 citations) CD45-Brilliant Violet 650 (3 citations) Anti-CD8α-Brilliant-Violet-650 (2 citations) CD4-Brilliant Violet 650 (2 citations) CD3 Brilliant Violet 650 (4 citations)

10 protocols using brilliant violet 650

1

Multiparametric Flow Cytometry Analysis

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The following flurochrome-conjugated antibodies were used for surface staining: anti-human CD3 (Brilliant Violet 650, 1:100, OKT3, BioLegend, San Diego, CA), CD4 (PeCy7, 1:100, SK3, BioLegend), CD8 (APC-Cy7, 1:100, SK1, BD Biosciences), CD19 (PE Texas Red, 1:100, SJ25-C1, Invitrogen or APC, HIB19, Biolegend), CD25 (FITC, 1:50, 2A3, BD Biosciences, San Jose, CA), CD31 (APC, 1:100, WM59, eBioscience, San Diego, CA), CD45RA (Brilliant Violet 605, 1:100, HI100, BioLegend), CD45RO (PerCpFl710, 1:100, UCHL1, eBioscience), CD69 (Brilliant Violet 421, 1:100, FN50, BioLegend or BUV395, 1:100, FN50, BD Biosciences), CD103, (Alexa Fluor 647, 1:100, Ber-ACT8, BioLegend), CD127 (BV711, 1:100, A019D5, BioLegend), CCR7 (Alexa Fluor 488, 1:100, TG8, BioLegend). For intracellular staining, surface stained cells were resuspended and incubated in fixation buffer (eBioscience), washed, resuspended in 0.1mL permeabilization buffer (eBioscience) and stained with anti-Foxp3 antibodies (PE, 1:20, 236A/E7, eBioscience) and Ki67 (α700, 1:100, Ki-67, BioLegend) for 30 min at room temperature and washed twice with permeabilization buffer. Stained cells were acquired on a 6-laser LSRII analytical flow cytometer (BD Biosciences) in the CCTI flow cytometry core and analyzed using FlowJo software (Treestar, Ashland, OR).
Thome J.J., Bickham K.L., Ohmura Y., Kubota M., Matsuoka N., Gordon C., Granot T., Griesemer A., Lerner H., Kato T, & Farber D.L. (2015). Early life compartmentalization of human T cell differentiation and regulatory function in mucosal and lymphoid tissues. Nature medicine, 22(1), 72-77.
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2

Flow Cytometry Labeling of HIV-1 Envelope Trimers

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Biotinylated BG505 SOSIP MD39 trimer and BG505 SOSIP.v5.2 N241/N289 trimers were generated as previously described (17 (link), 23 (link)). Biotinylated proteins were individually premixed with fluorochrome-conjugated streptavidin (Brilliant Violet 650 or Brilliant Violet 421, BioLegend) at room temperature for 20 min. BG505 SOSIP MD39-ferritin and BG505 SOSIP T33-31 nanoparticles were generated and directly conjugated to Alexa Fluor 647 (Thermo Fisher Scientific). Cells were incubated with indicated probes for 30 min at 4°C, and then surface antibodies were added and incubated for 30 min at 4°C. Cells were washed and then sorted on a BD FACSAria II. The sorting procedure is shown in fig. S6.
Antanasijevic A., Bowman C.A., Kirchdoerfer R.N., Cottrell C.A., Ozorowski G., Upadhyay A.A., Cirelli K.M., Carnathan D.G., Enemuo C.A., Sewall L.M., Nogal B., Zhao F., Groschel B., Schief W.R., Sok D., Silvestri G., Crotty S., Bosinger S.E, & Ward A.B. (2022). From structure to sequence: Antibody discovery using cryoEM. Science Advances, 8(3), eabk2039.
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3

Multicolor Flow Cytometry Panel

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Alexa Fluor® 647 anti-mouse IDO1 (2E2/IDO1), PE anti-mouse CD11c (N418), PerCP/Cyanine5.5 anti-mouse/human CD11b (M1/70), Brilliant Violet 650™ anti-mouse CD80 (16-10A1), APC anti-mouse CD86 (GL-1), FITC anti-mouse I-A/I-E (M5/114.15.2), PerCP/Cyanine5.5 anti-mouse CD45 (30-F11), APC/Fire™ 750-anti-mouse CD3 (17A2), FITC anti-mouse CD4 (RM4-5), Brilliant Violet 650™ anti-mouse CD8a (53-6.7), APC anti-mouse CD62L (MEL-14), PE/Dazzle™594 anti-mouse/human CD44 (IM7), PE/Dazzle™594-anti-mouse CD184 (CXCR4) (L276F12), Brilliant Violet 421™ anti-mouse CD197 (CCR7) (4B12), PE anti-human/mouse Granzyme B (QA16A02), Brilliant Violet 421™ anti-mouse IFN-γ (XMG1.2), Brilliant Violet 421™ anti-mouse Ki-67 (16A8) were purchased from Biolegend, USA.
Meng R., Fu Y., Zhang Y., Mou Y., Liu G, & Fan H. (2022). Indoleamine 2,3-dioxygenase 1 signaling orchestrates immune tolerance in Echinococcus multilocularis-infected mice. Frontiers in Immunology, 13, 1032280.
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4

Multiparametric Flow Cytometry Analysis

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Brilliant Violet 421™ anti-mouse I-A/I-E Antibody, Brilliant Violet 510™ anti-mouse CD4 Antibody, Brilliant Violet 570™ anti-mouse/human CD45R/B220 antibody, Brilliant Violet 605™ anti-mouse IgM Antibody, Brilliant Violet 650™ anti-mouse CD25 antibody, Brilliant Violet 785™ anti-mouse CD8a antibody, PE/Cy7 Goat anti-mouse IgG (minimal x-reactivity) antibody, Alexa Fluor® 488 anti-mouse/rat/human FOXP3 antibody, Alexa Fluor® 647 anti-mouse IgD antibody, and APC/Cy7 anti-mouse CD138 (Syndecan-1) antibody were purchased from Biolegend. AnaSpec 7-AAD was procured from Fisher Scientific and Biotinylated Peanut Agglutinin (PNA), was purchased from Vector Labs. Biotinylated goat anti-mouse IgG, IgG1, IgA and IgM secondary Abs were purchased from Jackson ImmunoResearch Laboratories. For monitoring protein expression on arrays, monoclonal mouse anti-polyhistidine was procured from Sigma-Aldrich (St. Louis, MO, USA) and rat anti-hemagglutinin (HA; clone 3F10, anti-HA high affinity), from Roche (Pleasanton, CA) were used. Streptavidin-conjugated SureLight P3 was purchased from Columbia Biosciences (Frederick, MD).
Dotsey E., Ushach I., Pone E., Nakajima R., Jasinskas A., Argueta D.A., Dillon A., DiPatrizio N., Davies H., Zlotnik A., Crompton P.D, & Felgner P.L. (2017). Transient Cannabinoid Receptor 2 Blockade during Immunization Heightens Intensity and Breadth of Antigen-specific Antibody Responses in Young and Aged mice. Scientific Reports, 7, 42584.
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5

Comprehensive Immune Profiling of PBMCs

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The PBMCs were collected from patient blood using human lymphocyte separation medium. A total of 0.5×106 to 1×106 PBMCs were washed by 1x PBS and incubated with the indicated antibodies for 30 minutes at 4°C. For intracellular staining, cells were washed, fixed and permeabilized using the transcription factor staining buffer set (eBiosciences, #00–5523–00). Antibodies for intracellular staining were incubated with cells for 20 minutes at 4°C. Cells were washed, resuspended in 1x PBS, and acquired on the cytoFLEX LX Flow Cytometer. Data were analyzed using FlowJo V.10. The antibodies that were used are listed as follows: CD3 (PerCP/Cyanine5.5, clone UCHT1, catalog No. 300430, BioLegend), CD4 (APC/Cyanine7, clone SK3, catalog No. 317418, BioLegend), CD56 (PE-Cy7, catalog No. 985912, BD Biosciences), PD-1 (Brilliant Violet 421, catalog No. 367421, BioLegend), PD-L1 (APC, catalog No. 329707, BioLegend), CD68 (APC, catalog No. 333809, BioLegend), CD86 (FITC, catalog No. 555657, BD Biosciences) and Foxp3 (Brilliant Violet 421, catalog No. 320123, BioLegend), CD163 (APC/Cyanine7, catalog No. 333621, BioLegend), HLA-DR (Brilliant Violet 650, catalog No. 307649, BioLegend), CD11c (PerCP/Cyanine5.5, catalog No. 980610, BioLegend), and Fixable Viability Dye eFluor 506 (eBiosciences, #65–0866–18).
Hong D.C., Yang J., Sun C., Liu Y.T., Shen L.J., Xu B.S., Que Y., Xia X, & Zhang X. (2023). Genomic Profiling of Radiation-Induced Sarcomas Reveals the Immunologic Characteristics and Its Response to Immune Checkpoint Blockade. Clinical Cancer Research, 29(15), 2869-2884.
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6

Apoptosis Induction in PBMC Subsets

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PBMCs from PROMOTE subjects (28 months of age; no chemoprevention control arm) were thawed and rested overnight either in standard media (untreated), 5uM camptothecin (Sigma) or P. falciparum schizont extract (PfSE) or protein extract from uninfected RBCs (uE) at an effector:target ratio of 3:1. To test for induction of apoptosis, stains for AnnexinV (Biolegend) or YoPro (Invitrogen), or activated Caspase 3 FITC (BD) were used according to the manufacturer’s instructions in combination with the following antibodies: AnnexinV and YoPro staining—CD3 (OKT3) Brilliant Violet 650, CD4 (RPA-T4) APC-Cy7, CD127 (A019D5) APC, CD25 (BC96) PE-Cy7 from Biolegend; for Caspase3—CD3 (OKT3) Brilliant Violet 650, CD4 (RPA-T4) PerCP, CD127 (A019D5) Pacific Blue, CD25 (BC96) PE-Cy7. Tregs were gated as CD3+CD4+CD25+CD127dim. Sensitivity to apoptosis was measured ex vivo (untreated control), after induction with camptothecin (fold change compared to untreated), and after stimulation with P. falciparum schizont extract (fold change comparing PfSE to uE).
Boyle M.J., Jagannathan P., Farrington L.A., Eccles-James I., Wamala S., McIntyre T.I., Vance H.M., Bowen K., Nankya F., Auma A., Nalubega M., Sikyomu E., Naluwu K., Rek J., Katureebe A., Bigira V., Kapisi J., Tappero J., Muhindo M.K., Greenhouse B., Arinaitwe E., Dorsey G., Kamya M.R, & Feeney M.E. (2015). Decline of FoxP3+ Regulatory CD4 T Cells in Peripheral Blood of Children Heavily Exposed to Malaria. PLoS Pathogens, 11(7), e1005041.
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7

Multicolor Flow Cytometry of Immune Cells

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The following antibodies were used for flow cytometry: Alexa Fluor488-anti-CD45RA, phycoerythrin (PE)–anti-CD57, allophycocyanin/cyanine 7 (Cy7) or PE/Cy7–anti-CD3, Alexa Fluor 647–anti-CX3CR1, Alexa Fluor 700–anti-CD27, Brilliant Violet 510–anti-CD28, PE/Cy7–anti-CD8 and Brilliant Violet 650–anti-CD279 (programmed cell death protein-1-PD-1) (all from Biolegend); LIVE/DEAD fixable violet dye (Molecular Probes); allophycocyanin–eFluor 780–anti-CD4 (both from ebiosciences)
Garg A., Gianella S., Nakazawa M., Trout R, & Spector S.A. (2019). Association of Cytomegalovirus DNA and Immunologic Markers of Cardiovascular Disease. Open Forum Infectious Diseases, 6(5), ofz113.
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8

Phenotyping of NK cell subsets

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KIR3DL1/S1 expression was detected on NK cells using the following monoclonal antibodies: anti-KIR3DL1 (clone:DX9, Brilliant Violet 421, Biolegend, San Diego, CA, USA), anti-KIR3DL1/S1 (clone:Z27, APC, Beckman Coulter, Brea, CA, USA), anti-CD56 (clone:N901, ECD, Beckman Coulter) and anti-CD3 (OKT3, Brilliant Violet 650, Biolegend). Dead cells were excluded by staining with DAPI and NK cells were classified as CD3-CD56+. All FACS analyses were performed on an LSR Fortessa (Beckton Dickenson, San Jose, CA, USA) and analyzed using FlowJo 9.7 software (Treestar, Ashland, OR, USA).
Boudreau J.E., Le Luduec J.B, & Hsu K.C. (2014). Development of a Novel Multiplex PCR Assay to Detect Functional Subtypes of KIR3DL1 Alleles. PLoS ONE, 9(6), e99543.
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9

Multiparametric Flow Cytometry Analysis

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The following flurochrome-conjugated antibodies were used for surface staining: anti-human CD3 (Brilliant Violet 650, 1:100, OKT3, BioLegend, San Diego, CA), CD4 (PeCy7, 1:100, SK3, BioLegend), CD8 (APC-Cy7, 1:100, SK1, BD Biosciences), CD19 (PE Texas Red, 1:100, SJ25-C1, Invitrogen or APC, HIB19, Biolegend), CD25 (FITC, 1:50, 2A3, BD Biosciences, San Jose, CA), CD31 (APC, 1:100, WM59, eBioscience, San Diego, CA), CD45RA (Brilliant Violet 605, 1:100, HI100, BioLegend), CD45RO (PerCpFl710, 1:100, UCHL1, eBioscience), CD69 (Brilliant Violet 421, 1:100, FN50, BioLegend or BUV395, 1:100, FN50, BD Biosciences), CD103, (Alexa Fluor 647, 1:100, Ber-ACT8, BioLegend), CD127 (BV711, 1:100, A019D5, BioLegend), CCR7 (Alexa Fluor 488, 1:100, TG8, BioLegend). For intracellular staining, surface stained cells were resuspended and incubated in fixation buffer (eBioscience), washed, resuspended in 0.1mL permeabilization buffer (eBioscience) and stained with anti-Foxp3 antibodies (PE, 1:20, 236A/E7, eBioscience) and Ki67 (α700, 1:100, Ki-67, BioLegend) for 30 min at room temperature and washed twice with permeabilization buffer. Stained cells were acquired on a 6-laser LSRII analytical flow cytometer (BD Biosciences) in the CCTI flow cytometry core and analyzed using FlowJo software (Treestar, Ashland, OR).
Thome J.J., Bickham K.L., Ohmura Y., Kubota M., Matsuoka N., Gordon C., Granot T., Griesemer A., Lerner H., Kato T, & Farber D.L. (2015). Early life compartmentalization of human T cell differentiation and regulatory function in mucosal and lymphoid tissues. Nature medicine, 22(1), 72-77.
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10

Multiparameter Flow Cytometry Analysis

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Phenotypic analyses of cells were performed using a BD FACSAria III Cytometer (BD Biosciences) at our Flow Cytometry Core Facility and the following human antibodies (all from Biolegend) were used: lineage negative (PE-conjugated anti-CD19, CD3, CD1a, CD11b, CD34, FcεRIα, CD14, CD11c, CD94), Brilliant Violet 650™ -conjugated anti-CD45, FITC-conjugated anti-CD56, Alexa Fluor® 647-conjuated anti-NKp44, PE/Cy7-conjuated anti-CD117, PerCP/Cyanine5.5 anti-CD294 (CRTH2) and Brilliant Violet 421™ -conjugated anti-CD127. Cells were then washed with cell staining buffer (Biolegend) for removing excess antibody and resuspended in 4% paraformaldehyde solution and incubated at room temperature for 20 minutes. Data were analyzed using FlowJo software (Tree Star, Ashland, OR, USA). The gating strategy utilized appropriate fluorescent minus one (FMO) negative controls. Any samples with viability of 30% or lower (as determined by staining with live dead marker Zombie NIR™, BioLegend) were excluded from all analyses. Any samples with lower than 25% CD45+ were excluded from all analyses.
Kang J., Loh K., Belyayev L., Cha P., Sadat M., Khan K., Gusev Y., Bhuvaneshwar K., Ressom H., Moturi S., Kaiser J., Hawksworth J., Robson S.C., Matsumoto C.S., Zasloff M., Fishbein T.M, & Kroemer A. (2020). Type 3 innate lymphoid cells are associated with a successful intestinal transplant. American journal of transplantation : official journal of the American Society of Transplantation and the American Society of Transplant Surgeons, 21(2), 787-797.
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