Cd158e1 bv421

Manufactured by BioLegend

CD158e1 BV421 is a fluorescently-labeled monoclonal antibody that recognizes the CD158e1 cell surface antigen. CD158e1 is a member of the killer cell immunoglobulin-like receptor (KIR) family and is expressed on natural killer (NK) cells and a subset of T cells. The BV421 fluorescent dye is conjugated to the antibody, allowing for its detection and quantification using flow cytometry.

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Lab products found in correlation

Lsrii flow cytometer, by BD (1 mentions) Cd56 pe cy7, by BioLegend (1 mentions) Cd107a fitc, by BD (1 mentions) Cd3 ecd, by Beckman Coulter (1 mentions) Apc cy7, by BioLegend (1 mentions) Tnf α af647, by BioLegend (1 mentions) Cd158b fitc, by BD (1 mentions) Navios flow cytometer, by Beckman Coulter (1 mentions) Cd161 apc, by Miltenyi Biotec (1 mentions) Cd56 ecd, by Beckman Coulter (1 mentions) Cd133 apc, by Miltenyi Biotec (1 mentions) Lsrfortessa cell analyzer, by BD (1 mentions) Nkg2d apc, by Miltenyi Biotec (1 mentions) Cd16 ecd, by Beckman Coulter (1 mentions) Nkp30 pe, by BioLegend (1 mentions) Nkp44 pe, by Beckman Coulter (1 mentions) Cd57 bv421, by BD (1 mentions) Nkp46 pe cy7, by BioLegend (1 mentions)

3 protocols using cd158e1 bv421

Flow Cytometric Analysis of NK Cell Receptors

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The antibodies used in this study were CD56 PE-Cy7 and APC-Cy7, CD158a/CD158b/CD158e1 PE (used in experiments were KIR were pooled), CD158e1 BV421, TNF-α AF647, IFN-γ Pacific blue, DLL1 and DLL4 PE, purified mouse IgM isotype control (BioLegend), CD158b FITC, CD107a FITC, purified mouse anti-human CD16 (BD Bioscience), CD3 ECD, CD158b APC (Beckman Coulter), and CD158a PE-Cy7 (eBioscience). For CD16 activation studies, cells were cultured with anti-CD16 or isotype control for 30 min and then crosslinked with goat-anti mouse IgG for 5 hours. Staining, acquisition, and analysis were performed as previously described (27 (link)). Finally, cells were run on an LSRII flow cytometer (BD Biosciences) and data were analyzed with FlowJo software (TreeStar).

Felices M., Ankarlo D.E., Lenvik T.R., Nelson H.H., Blazar B.R., Verneris M.R, & Miller J.S. (2014). Notch signaling at later stages of Natural Killer cell development enhances KIR expression and functional maturation. Journal of immunology (Baltimore, Md. : 1950), 193(7), 3344-3354.

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Phenotypic Characterization of RSV-Infected NK Cells

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The following fluorochrome-conjugated monoclonal antibodies were used to phenotypically characterize (RSV-infected) NK cells: CD3-APCAF750 (UCHT1, Beckman Coulter), CD16-PacificOrange (3G8, Thermo Fisher), CD56-ECD (N901, Beckman Coulter), CD85j-PerCP-Cy5.5 (ILT2, LILRB1; GHI/75, BioLegend), CD161-APC (191B8, Miltenyi), CD158a-AF700 (KIR2DL1; 143211, R&D Systems), CD158a/h-PC5.5 (KIR2DL1/S1; EB6B, Beckman Coulter), CD158b1/b2,j-PC7 (KIR2DL2/L3/S2; GL183, Beckman Coulter), CD158e1-BV421 (KIR3DL1; DX9, BioLegend), CD159a-APC (NKG2A; Z199, Beckman Coulter), CD159c-PE (NKG2C; 134591, R&D Systems), CD244-AF700 (2B4; C1.7, BioLegend), CD314-APC (NKG2D; ON72, Beckman Coulter), CD335-PC7 (NKp46; BAB281, Beckman Coulter), CD336-PE (NKp44; Z231, Beckman Coulter), CD337-PerCP-Cy5.5 (NKp30; P30-15, BioLegend), RSV-G-FITC (131-2G, Millipore). Cells were measured using a Navios flow cytometer (Beckman Coulter).

van Erp E.A., Feyaerts D., Duijst M., Mulder H.L., Wicht O., Luytjes W., Ferwerda G, & van Kasteren P.B. (2018). Respiratory Syncytial Virus Infects Primary Neonatal and Adult Natural Killer Cells and Affects Their Antiviral Effector Function. The Journal of Infectious Diseases, 219(5), 723-733.

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NK Cell Surface Marker Profiling

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NK3.3-LTV cells were analyzed for changes in cell surface marker expression compared to non-immortalized NK3.3 cells. All cells (1 x 10⁵ cells per antibody) were centrifuged at 300 g x 5 min, media aspirated, and 1.5 µL single antibody + 20 µL staining buffer (phosphate buffered saline (PBS) + 1% bovine serum albumin + 1% sodium azide) were added to cells. Sample tubes were incubated for 30 min on ice, followed by addition of 200 µL staining buffer and incubated for another 10 min on ice. Samples were analyzed using LSRFortessa Cell Analyzer (BD Biosciences, Franklin Lakes, NJ). Data were analyzed using FlowJo software. Antibodies used include: NKp46 PE/Cy7 (#331915, BioLegend, San Diego, CA); NKp30 PE (#325207, BioLegend); NKp44 PE (#IM3710, Beckman Coulter, Brea, CA); CD94 PE (#130-098-974, Miltenyi Biotec, Auburn, CA); NKG2D APC (#120-003-706, Miltenyi Biotec); CD161 APC-Alexa Fluor750 (#B30630, Beckman Coulter); CD158e1 BV421 (#312713, BioLegend); CD158a,h PE/Cy7 (#A66899, Beckman Coulter); CD244 PerCp Cy5.5 (#B21171, Beckman Coulter); CD16 ECD (#A33098, Beckman Coulter); CD133 APC (#130-098-829, Miltenyi Biotec); CD57 BV421 (#563896, BD Biosciences); CD158 FITC (#339503, BioLegend).

Matchett E.C, & Kornbluth J. (2023). Extracellular vesicles derived from immortalized human natural killer cell line NK3.3 as a novel therapeutic for multiple myeloma. Frontiers in Immunology, 14, 1265101.

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