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Anti nk1.1 apc cy7

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Anti-NK1.1 APC-cy7 is a fluorescent-conjugated antibody that binds to the NK1.1 antigen, which is expressed on natural killer (NK) cells and a subset of T cells in mice. This antibody can be used for the identification and enumeration of NK1.1-positive cells in flow cytometric analysis.

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Lab products found in correlation

Anti cd8 fitc, by BioLegend (2 mentions) Anti cd45 pacific blue, by BioLegend (2 mentions) Anti cd4 pe, by BioLegend (2 mentions) Anti ter119 fitc, by BioLegend (2 mentions) Anti sca1 pacific blue, by BioLegend (2 mentions) Anti pdgfrα pe, by BioLegend (2 mentions) Anti b220 apc cy7, by BioLegend (2 mentions) Ack solution, by Thermo Fisher Scientific (2 mentions) Anti cd45 fitc, by BioLegend (2 mentions) Anti cd31 fitc, by BioLegend (2 mentions) Anti cd3 apc, by BioLegend (2 mentions) Facsaria 2, by BD (2 mentions) Countbright absolute counting beads, by Thermo Fisher Scientific (1 mentions) Anti cd11b apc, by Thermo Fisher Scientific (1 mentions) Fitc anti gr 1, by BD (1 mentions) Anti cd4 pb, by BD (1 mentions) Anti cd69 pe, by BioLegend (1 mentions) Anti cd3 alexa 700, by BD (1 mentions) Anti b220 percp, by BD (1 mentions) Anti cd11c pecy7, by Thermo Fisher Scientific (1 mentions)

4 protocols using anti nk1.1 apc cy7

1

Multicolor Flow Cytometry of Tumors

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Cells were analyzed using LSRII (BD, Pharmingen) and sorted using FACS-ARIA II (BD, Pharmingen). The following antibodies were used: anti-CD45 FITC, anti-CD31 FITC, anti-Ter119 FITC, anti-Sca1 Pacific Blue, anti-PDGFRα PE, anti-CD3 APC, anti-CD8 FITC, anti-CD45 Pacific Blue, anti-CD4 PE, anti-NK1.1 APC-cy7, anti-B220 APC-cy7 (all purchased from Biolegend). Tumors were digested to single cell suspension enzymatically and filtered twice through 70 μm filters. Red blood cells were lysed with ACK solution (Gibco), washed twice with PBS, and then stained with the fluorophore-conjugated antibodies for 15 min at room temperature. The excess of unbound antibodies was washed out before acquisition in flow cytometry.
Tessaro F.H., Ko E.Y., De Simone M., Piras R., Broz M.T., Goodridge H.S., Balzer B., Shiao S.L, & Guarnerio J. (2022). Single-cell RNA-seq of a soft-tissue sarcoma model reveals the critical role of tumor-expressed MIF in shaping macrophage heterogeneity. Cell reports, 39(12), 110977.
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2

Splenic Immune Cell Profiling

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At the time of sacrifice, spleens were collected and single-cell suspensions were prepared from each animal. Samples were stained with anti-CD3-Alexa 700, anti-CD4-PB, anti-Gr-1-FITC, anti-B220-PerCP (BD Bioscience, San Jose, CA), anti-CD8-PO (Life Technologies, Carlsbad, CA), anti-NK1.1-APC-Cy7, anti-CD69-PE (Biolegend, San Diego, CA), anti-CD11b-APC, and anti-CD11c-PE-Cy7 (eBioscience, San Diego, CA). CountBright absolute counting beads (Life Technologies) were added to cell suspensions and were used according to manufacturer’s instructions to determine absolute cell counts. Samples were run on an LSR II flow cytometer (BD Biosciences). Resulting flow cytometry standard files were analyzed using FlowJo software (version 10.0.7, TreeStar, Ashland, OR).
Klingensmith N.J., Chen C.W., Liang Z., Burd E.M., Farris A.B., Arbiser J.L., Ford M.L, & Coopersmith C.M. (2018). Honokiol increases CD4+ T cell activation and decreases TNF but fails to improve survival following sepsis. Shock (Augusta, Ga.), 50(2), 178-186.
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3

Multicolor Flow Cytometry of Tumors

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Cells were analyzed using LSRII (BD, Pharmingen) and sorted using FACS-ARIA II (BD, Pharmingen). The following antibodies were used: anti-CD45 FITC, anti-CD31 FITC, anti-Ter119 FITC, anti-Sca1 Pacific Blue, anti-PDGFRα PE, anti-CD3 APC, anti-CD8 FITC, anti-CD45 Pacific Blue, anti-CD4 PE, anti-NK1.1 APC-cy7, anti-B220 APC-cy7 (all purchased from Biolegend). Tumors were digested to single cell suspension enzymatically and filtered twice through 70 μm filters. Red blood cells were lysed with ACK solution (Gibco), washed twice with PBS, and then stained with the fluorophore-conjugated antibodies for 15 min at room temperature. The excess of unbound antibodies was washed out before acquisition in flow cytometry.
Tessaro F.H., Ko E.Y., De Simone M., Piras R., Broz M.T., Goodridge H.S., Balzer B., Shiao S.L, & Guarnerio J. (2022). Single-cell RNA-seq of a soft-tissue sarcoma model reveals the critical role of tumor-expressed MIF in shaping macrophage heterogeneity. Cell reports, 39(12), 110977.
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4

Single-cell Tetramer Staining Protocol

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Single-cell suspensions were stained with PE or APC-labelled IAb/NP311-325 or APC-labelled Db/NP368-374 tetramers (NIH tetramer core) at 37°C, 5% CO2 for 2 h in complete RPMI (RPMI with 10% foetal calf serum, 100 μg/ml penicillin-streptomycin, and 2 mM l-glutamine) containing Fc block (24G2). Surface antibodies were added and the cells were incubated for a further 20 min at 4°C. Antibodies used were: anti-CD3 BV785 (BioLegend; clone: 17A2), anti-CD4 BUV805 (BD Bioscience; clone: RM4-5), anti-CD8 BV421 (ThermoFisher; clone: 53-6.7), anti-CD44 BUV395 (BD; clone: IM7), anti-CD45.2 BV605 (BioLegend; clone: 104), anti-CD69 PE-Cy7 (ThermoFisher; clone: H1.2F3), anti-CD127 APC (ThermoFisher; clone: A7R34), anti- γδ TCR PE-Cy7 (BioLegend; clone: GL3), anti-ICOS BV785 (BioLegend; clone: C398.4A), anti-NK1.1 APC-Cy7 (BioLegend; clone: PK136), anti-PD1 BV711 (BioLegend; clone: 29F,1A12), and ‘dump’ antibodies: B220 (RA3-6B2), F4/80 (BM8), and MHC II (M5114) all on eFluor-450 (ThermoFisher) or PerCP-Cy5.5 (ThermoFisher; B220 and F4/80, and BioLegend; MHCII). Cells were stained with a fixable viability dye eFluor 506 (ThermoFisher). Stained cells were acquired on a BD LSR Fortessa and analysed using FlowJo (version 10, BD Bioscience).
Finney G.E., Hargrave K.E., Pingen M., Purnell T., Todd D., MacDonald F., Worrell J.C, & MacLeod M.K. (2023). Triphasic production of IFNγ by innate and adaptive lymphocytes following influenza A virus infection. Discovery Immunology, 2(1), kyad014.
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