Cd56 brilliant violet 421

Manufactured by Sony

Sourced in United States

The CD56-Brilliant Violet 421 is a fluorescent dye used for flow cytometry analysis. It is designed to bind specifically to the CD56 antigen, which is expressed on natural killer cells and some subsets of T cells. The dye is excited by a violet laser and emits fluorescence in the Brilliant Violet 421 spectrum, allowing for the detection and quantification of CD56-positive cells in a sample.

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Lab products found in correlation

Cd3 pe cy7, by Beckman Coulter (2 mentions) Cd56 apc, by Miltenyi Biotec (1 mentions) Cd107a pe cy5, by Thermo Fisher Scientific (1 mentions) Cd2 pe cy7, by Sony (1 mentions) Hla dr pe, by Sony (1 mentions) Cd57 fitc clone tb03, by Miltenyi Biotec (1 mentions) Cd56 pe, by Agilent Technologies (1 mentions) Facscalibur flow cytometer, by BD (1 mentions) Cd56 pe, by Beckman Coulter (1 mentions) Cd56 apc, by Beckman Coulter (1 mentions) Cd16 pe, by Sony (1 mentions) Anti hla dr fitc, by Beckman Coulter (1 mentions) Cd57 pe, by Thermo Fisher Scientific (1 mentions) Cd57 fitc, by Miltenyi Biotec (1 mentions) Cd57 apc, by Miltenyi Biotec (1 mentions) Anti nkg2a pe, by R&D Systems (1 mentions) Facsvantage diva machine, by BD (1 mentions)

3 protocols using cd56 brilliant violet 421

Multiparameter Flow Cytometry for NK Cells

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For surface and intracellular staining of NK cells the following mouse anti-human fluorescent-labeled Abs were used: CD2-PE-Cy7 (clone TS1/8), CD56-BrilliantViolet421 (clone HCD56), CD56-AF488 (clone 5.1H11), CD56-PE-Cy7 (clone 5.1H11), CD57-PE (clone TB01), HLA-DR-FITC (clone LN3), HLA-DR-PE (clone L243), IFN-gamma-FITC (clone 4S.B3), KI-67-PE (clone Ki-67), NKp46-FITC (clone 9E2) (Sony Biotechnology, USA), CD16-PE (Sorbent, Russia), CD16-APC (clone REA423), CD56-APC (clone REA196), CD57-FITC (clone TB03), CD57-APC (clone TB03), CD57-PE-Vio770 (clone TB03) (Miltenyi Biotec, Germany), CD44-F (clone J.173, Immunotech, USA), CD56-PE (clone C5.9, Dako, USA), CD107a-PE-Cy5 (clone H4A3), NKp30-PE (clone P30-15) (eBioscience, USA), CD161-AlexaFluor647 (clone HP-3G10), Granzyme B-Alexa Fluor 647 (clone GB11), KIR2DL2/DL3-PE (clone DX27) (Biolegend, USA), FcεRIγ-FITC (polyclonal, Milli-Mark, Merck, Germany), NKG2A-PE (clone 131411), NKG2C-AlexaFluor 488 (clone 108724), NKG2C-PE (clone 134591) (R&D Systems, USA). Supernatant of hybridoma producing Abs to human KIR2DL1 was kindly provided by prof. Miguel Lopez-Botet (Universitat Pompeu Fabra, Barcelona, Spain). Sheep anti-mouse IgG-FITC and PE (Sigma-Aldrich, USA) were used as second Abs for anti-KIR2DL1. IgG1-PE (clone IS5-21F5, Miltenyi Biotec, Germany) was used as isotype control for NKG2C-PE Ab staining.

Kobyzeva P.A., Streltsova M.A., Erokhina S.A., Kanevskiy L.M., Telford W.G., Sapozhnikov A.M, & Kovalenko E.I. (2020). CD56dimCD57−NKG2C+ NK cells retaining proliferative potential are possible precursors of CD57+NKG2C+ memory-like NK cells. Journal of leukocyte biology, 108(4), 1379-1395.

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Phenotyping of Natural Killer Cells

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The following anti-human monoclonal antibodies (mAbs) were used for flow cytometry: CD3- PE-Cy7 (Beckman Coulter, USA, clone UCHT1), CD56-APC (Beckman Coulter, USA, clone N901), CD56-Brilliant Violet 421 (Sony, USA, clone HCD56), CD56-PE (Beckman Coulter, USA, clone N901 (HLDA6)), CD57-PE (eBioscience, USA, clone TB01), CD57-FITC (Miltenyi Biotech, Germany, clone TB03), CD57-APC (Miltenyi Biotech, Germany, TB03), CD16-PE (Sony, USA) anti-NKG2A-PE (R&D Systems, USA, clone 131411), anti-KIR2DL2/DL3-PE (Miltenyi Biotech, Germany, clone DX27), anti-HLA-DR-FITC (Beckman Coulter, USA, clone B8.12.2). Cells were incubated in PBS containing 0.5% BSA (bovine serum albumin) and 0.01% sodium azide (labeling buffer) with mAbs for 30 min on ice, then washed twice with labeling buffer and centrifugation. Samples were subsequently analyzed using FACSCalibur flow cytometer (BD Biosciences, San Jose CA, USA) equipped with 488 and 640 nm lasers. At least 30000 events were recorded in lymphocyte gate for total NK cell population and 5000 events for NK cell clones. Acquired data was analyzed using Flowing Software version 2.5.1 (PerttuTerho, Turku Centre for Biotechnology, Finland) and FlowJo software version 7.6 (FlowJo LLC, Ashland, OR, USA). For fluorescence-activated cell sorting, cells were labeled with mAbs in PBS containing 0.5% BSA and 2 mM EDTA.

Streltsova M.A., Erokhina S.A., Kanevskiy L.M., Lee D.A., Telford W.G., Sapozhnikov A.M, & Kovalenko E.I. (2018). Analysis of NK cell clones obtained using interleukin-2 and gene-modified K562 cells revealed the ability of “senescent” NK cells to lose CD57 expression and start expressing NKG2A. PLoS ONE, 13(12), e0208469.

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Expansion of Single NK Cell Clones

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Purified NK cells were labeled with mouse anti-human monoclonal antibodies (mAbs) CD3-PE-Cy7 (Beckman Coulter, Brea, CA, USA, clone UCHT1) and CD56-Brilliant Violet 421 (Sony Biotechnology Inc., San Jose, CA, USA, clone HCD56). CD3⁻CD56⁺ cells were sorted into 96-well round-bottom plates containing feeder cells suspended in complete NK cell medium, one cell per well in the “single cell” mode. Cell sorting was performed with FACSVantage DiVa machine (BD Biosciences, San Jose, CA, USA) equipped with 405, 488, and 643 nm lasers, as well as an appropriate set of detectors and filters. The plates were then put into a CO₂-incubator (5% CO₂, 37 °C) for two weeks. After eight weeks of incubation, clones were transferred to a new 96-well round-bottom plate.

Streltsova M.A., Erokhina S.A., Kanevskiy L.M., Grechikhina M.V., Kobyzeva P.A., Lee D.A., Telford W.G., Sapozhnikov A.M, & Kovalenko E.I. (2019). Recurrent Stimulation of Natural Killer Cell Clones with K562 Expressing Membrane-Bound Interleukin-21 Affects Their Phenotype, Interferon-γ Production, and Lifespan. International Journal of Molecular Sciences, 20(2), 443.

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