Flow cytometry was performed on isolated liver and spleen leukocytes from control and TAA-treated mice. Cells were co-stained with antibodies against CD3ɛ - PE/Cy7 (145-2C11, BioLegend, San Diego, USA), CD4-Pacific orange (RM4-5, Invitrogen), CD8α –Pacific blue (53–6.7, BioLegend), CD19 –AlexaFlour 488 (6D5, BioLegend), NK1.1-PE (PK136, BD Biosciences, San Jose, USA), F4/80-PerCP/Cy5.5 (RM8, BioLegend) and CD147 –biotin (OX114, BioLegend) further stained with Streptavidin-APC (Invitrogen). Flow cytometry data were collected with FACS LSR-II (BD Biosciences). Each leukocyte subset was analysed for expression of CD147 and data recorded as median fluorescence intensity. Bar graphs represent mean ± SEM. Statistical analysis was performed by the Kruskal-Wallis test followed by Dunn's multiple comparisons test.
Nk1.1 pe pk136
Manufactured by BD
Sourced in United States
NK1.1-PE (PK136) is a fluorescently labeled antibody that binds to the NK1.1 antigen, which is expressed on natural killer cells and a subset of T cells in mice. It is commonly used in flow cytometry applications to identify and characterize these cell populations.
Automatically generated - may contain errors
Lab products found in correlation
Lsrii flow cytometer, by BD (2 mentions)
Live dead fixable aqua dead cell stain kit, by Thermo Fisher Scientific (2 mentions)
Facs lsr 2, by BD (1 mentions)
Streptavidin apc, by Thermo Fisher Scientific (1 mentions)
Cd11b apc m1 70, by BD (1 mentions)
Ly6g apc cy7 1a8, by BD (1 mentions)
Lrsiii hts, by BD (1 mentions)
Cell strainer, by BD (1 mentions)
Ter119 pe, by BD (1 mentions)
Cd45r pe, by Thermo Fisher Scientific (1 mentions)
Facsaria 3, by BD (1 mentions)
Pe cd11b m1 70, by Thermo Fisher Scientific (1 mentions)
40 m cell strainer, by BD (1 mentions)
Sytox blue, by Thermo Fisher Scientific (1 mentions)
Percp cy5.5 ly6c, by BD (1 mentions)
Fixation permeabilization solution kit, by BD (1 mentions)
5 protocols using nk1.1 pe pk136
1
Flow Cytometry Analysis of Immune Cells in Liver and Spleen
2
Infection and Flow Cytometry Analysis
3
Bone Marrow Cell Isolation and Transplantation
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PanRosaYFP mice were used as donors. Bone marrow cells of PanRosaYFP mice were flushed from femurs, tibias, coxae, and humeri using PBS supplemented with 5% heat-inactivated FCS. Cells were filtered through a 40-µm cell strainer (Falcon). Fc receptors were blocked by incubating cells in 5% FCS with purified mouse IgG (500 mg/ml; Jackson ImmunoResearch). All stainings were performed in 5% FCS on ice for 30 min with optimal dilutions of commercially prepared antibodies. Reagents used were CD3ε PE (145-C11), CD11b PE (M1/70), CD45R PE (RA3-6B2), CD117 eFluor780 (2B8), Sca-1 PerCP-Cy5.5 (D7; eBiosciences), CD4 PE (H129.19), CD8a PE (53–6.7), CD19 PE (1D3), Gr-1 PE (RB6-8C5), NK1.1 PE (PK136), and Ter119 PE (Ter119; BD Pharmingen). The lineage cocktail was composed of CD3ε, CD4, CD8a, CD11b, CD19, CD45R, Gr-1, NK1.1, and Ter119. Dead cells were excluded by staining with Sytox Blue (Invitrogen). Approximately 5,000 LSK cells were sorted by FACSAriaIII (Becton Dickinson) and injected intravenously into nonirradiated triple transgenic Rag2−/−γc−/−KitW/Wv recipient mice. Donor chimerism of blood cells was determined 1 mo after transplantation.
4
Multiparametric Flow Cytometry Profiling
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Cells were washed twice with Dulbecco’s PBS containing 0.5% bovine serum albumin and 0.05% sodium azide. After washing, the cells were stained with LIVE/DEADTM Fixable Aqua Dead Cell Stain Kit (Invitrogen, Eugene, OR) following the manufacturer’s protocol. The cells were stained with fluorescence-conjugated surface antibodies for 30 min at 4 °C, washed, and analyzed using LSR II flow cytometer (BD Bioscience, San Jose, CA). The data were analyzed using FlowJo software (TreeStar, Ashland, OR). Antibodies used for flow cytometric analyses included: APC-Cy7 CD11b (M1/70), APC CD11b (M1/70), PerCP-Cy5.5 Ly6G (1A8), FITC Ly6C (AL21), PerCP-Cy5.5 Ly6C (AL-21), PerCP-Cy5.5 CD45.1 (A20), PE-Cy7 CD3e (145-2C11) PerCP-Cy5.5 CD4 (RM4-5), APC CD8a (53-6.7), and PE NK1.1 (PK136) (all from BD Bioscience); PE-Cy7 F4/80 (BM8), PE F4/80 (BM8) eFluor® 450 CD45 (30-F11) (all from eBioscience, San Diego, CA); PE CX3CR1 (polyclonal) and APC CCR2 (#475301) (both from R&D Systems, Madison, WI) for mouse. BV421 CD45 (HI30), PE CD14 (MφP9), and PerCP-Cy5.5 CD16 (3G8) (all from BD Bioscience); FITC CD68 (Y1/82A), APC CX3CR1 (FN50), and APC-Cy7 CCR2 (REA264) (all from eBioscience) and APC CD206 (19.2) (Biolegend, San diego, CA) for human.
5
Multiparametric Flow Cytometry of Liver Immune Cells
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Liver MNCs were washed with PBS and suspended in PBS containing 0.5% bovine serum albumin and 0.05% sodium azide. The cells were stained with LIVE/DEAD™ Fixable Aqua Dead Cell Stain Kit (Invitrogen) following the manufacturer's protocol. After washed, the cells were stained with fluorescence-conjugated surface antibodies for 30 min at 4°C. For intracellular cytokine staining, cells were fixed and permeabilized with Fixation/Permeabilization Solution Kit (BD Bioscience, San Jose, CA, USA). Permeabilized cells were stained with fluorescence-conjugated surface antibodies for 30 min at 4°C. Stained cells were analyzed using LSR II flow cytometer (BD Bioscience). The data were analyzed using FlowJo software (TreeStar, Ashland, OR, USA). Antibodies for flow cytometric analysis included: FITC CD3e (145-2C11), APC CD8a (53-6.7), APC-Cy7 CD11b (M1/70), PE Gr1 (RB6-8C5), PE IL-17A (TC11-18H10), FITC CD4 (RPA-T4), APC IL-10 (JES5-16E3), and PE NK1.1 (PK136) (all from BD Bioscience); eFluor® 450 CD45 (30-F11), FITC F4/80 (BM8), APC Foxp3 (FJK-16s), and PE CD25 (PC61.5) (all from eBioscience, San Diego).
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