Cd94 fitc

Manufactured by Thermo Fisher Scientific

The CD94 FITC is a monoclonal antibody that binds to the CD94 cell surface receptor. CD94 is a member of the C-type lectin receptor family and is expressed on natural killer (NK) cells, a subset of T cells, and other immune cells. The FITC (Fluorescein Isothiocyanate) label allows for the detection and identification of CD94-positive cells using flow cytometry or other fluorescence-based techniques.

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3 protocols using cd94 fitc

Differential Immunophenotyping of NK Cell Subsets

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The following antibodies were used. From eBioscience (San Diego, CA): CD3-APC eFluor 780 (clone SK7); CD16-FITC (eBioCB16); CD19-APC eFluor 780 (HIB19); CD45-PE (HI30); CD94-FITC (DX22); Eomes-PE eFluor 610 (WD1928); Granzyme K-PerCP eFluor 710 (G3H69); HLA-A3-FITC (GAP.A3); IFN gamma-Alexa Fluor 488 (4S.B3); S1PR1-eFluor 660 (SW4GYPP); T-bet-PECy7 (4B10); TNF alpha-APC (Mab11). From Biolegend (London, UK): CCR5-APC (J418F1); CD49a-FITC (TS2/7); CD69-APC (FN50); CD103-FITC (Ber-ACT8); CX3CR1-FITC (2A9-1); CXCR6-PerCP Cy5.5 (K041E5); CXCR6-APC (K041E5); GM-CSF-PE (BDV-21C11); Granzyme B-FITC (GB11); HLA-A2-FITC (BB7.2); KIR2DL1/S1/S3/S5-APC (HP-MA4); KIR2DL2/L3-APC (DX27); KIR3DL1-APC (DX9); Perforin-APC (dG9). From BD (Oxford, UK): CD56-BV510 (NCAM16.2), LIF-PE (1F10). Dead cells were excluded using Fixable Viability Dye eFluor 450 (eBioscience). Intracellular staining was carried out using Human FoxP3 Buffer (BD) according to the manufacturer’s instructions. Data were acquired on a Fortessa II (BD) and analysed using FlowJo (Treestar, Ashland, OR). Cells were sorted on an Aria (BD). Eomes^lo NK cells were isolated by sorting on live cells (propidium iodide negative, Tonbo Biosciences, San Diego, CA), singlets, scatter, CD3^- CD56⁺ CXCR6^- CD16⁺. Eomes^hi NK cells were isolated by sorting on live cells, singlets, scatter, CD3^- CD56⁺ CXCR6⁺.

Cuff A.O., Robertson F.P., Stegmann K.A., Pallett L.J., Maini M.K., Davidson B.R, & Male V. (2016). Eomeshi NK Cells in Human Liver Are Long-Lived and Do Not Recirculate but Can Be Replenished from the Circulation. The Journal of Immunology Author Choice, 197(11), 4283-4291.

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Comprehensive Immune Cell Phenotyping

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Antibodies used were as follows: CD56 BV421, CD3 PE-Cy7 or APC-Cy7, CD14 APC-Cy7, CD19 APC-Cy7, CD57 Pacific Blue or FITC, CD62L PE-Cy7, CD107a PE-Cy7, CD244 PE-Cy5.5, Perforin Pacific blue, CD160 Alexa Fluor 647 (Biolegend), CD16 eFluor450 or FITC, CD69 FITC, CD94 FITC, CD8a PerCP-Cy5.5 (eBioscience), CD161 PE or APC, CD56 APC, IFNγ FITC, NKp30 APC, NKp46 APC, NKp80 APC (Miltenyi Biotec), CD3 Pacific Orange, CD4 Qdot 605, Granzyme B APC (Invitrogen), NKG2C Alexa Fluor 488 or PE, NKG2D PE, PLZF APC, Granzyme A FITC (R&D Systems), CD56 FITC or PE-Cy7, CD85j FITC, IFNγ Alexa Fluor 700, Ki67 FITC (BD Biosciences), Granzyme K FITC (Immunotools), NKG2A PE, NKp44 PE, CD158e1/e2 PE (Beckman Coulter), Phosphatidylserine Alexa Fluor 488 (Merck Milipore). The viability dye Live/Dead fixable Near-IR (Invitrogen) was used in all experiments. Anti-KLRG1 FITC was kindly provided by H. Pircher. Data were acquired on LSRII (BD Biosciences) and analyzed using FlowJo (Treestar, Inc.).

Kurioka A., Cosgrove C., Simoni Y., van Wilgenburg B., Geremia A., Björkander S., Sverremark-Ekström E., Thurnheer C., Günthard H.F., Khanna N., Walker L.J., Arancibia-Cárcamo C.V., Newell E.W., Willberg C.B, & Klenerman P. (2018). CD161 Defines a Functionally Distinct Subset of Pro-Inflammatory Natural Killer Cells. Frontiers in Immunology, 9, 486.

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Comprehensive Murine NK Cell Immunophenotyping

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The following antibodies were purchased from Biolegend: CD3 BV510, CD49b (DX5) APC-Cy7, NKp46 BV421, CD11b PE, Ly49D PE, Streptavidin BV650, CD27 biotin, KLRG1 BV605, KLRG1 PerCP/Cy5.5, CD69 PerCP/Cy5.5, and Ki67 APC. The following antibodies were purchased from eBioscience: CD107a PerCP-eFluor710, Ly49I PE, CD94 FITC, IFNγ APC, and Ly49H PE-Cyanine7. The following antibodies were purchased from BD Biosciences: NKG2A/C/E BV605. The following antibodies were purchased from Miltenyl Biotec: NKG2D (CD314) biotin, Ly49D biotin, and Ly49D PerCP-Vio700. To determine viability Invitrogen Fixable live/dead Aqua stain was used.

Ivanova D.L., Fatima R, & Gigley J.P. (2016). Comparative Analysis of Conventional Natural Killer Cell Responses to Acute Infection with Toxoplasma gondii Strains of Different Virulence. Frontiers in Immunology, 7, 347.

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