Flow cytometry was utilized to determine cell phenotype and cell proliferation. Cells were stained for surface markers using fluorochrome conjugated antibodies for 30 minutes on ice, washed, acquired using FACSCantoII (BD Biosciences) and analyzed using FlowJo v10.4.2 (Tree Star LLC). Antibodies and their specific clones used were: CD3-FITC (17A2, eBioscience), CD3-PerCPCy5.5 (145–2C11, Tonbo Biosciences), CD4-Pacific Blue (RM4–5, eBioscience), CD4-PerCPCy5.5 (RM4–5, Tonbo Biosciences), CD8α-APC-Cy7 (53–6.7, eBioscience), CD8α-eFluor450 (53–6.7, eBioscience), CD11b-PercpCy5.5 (M1/70, Tonbo Biosciences), CD11c-APC-eFluor780 (N418, eBioscience), B220-PECy7 (RA3–6B2, eBioscience), CD44-PECy7(IM7, eBioscience), Foxp3-FITC (FJK-16s, eBioscience), I-Ab-APC(AF6–120.1, eBioscience), interferon-γ-APC (XMG1.2, eBioscience), violet proliferation dye (VPD450, BD Biosciences), and aqua live/dead dye (Molecular Probes).
Cd11b percpcy5.5 m1 70
Manufactured by Cytek Biosciences
The CD11b-PercpCy5.5 (M1/70) is a fluorescently-labeled monoclonal antibody used for the detection and analysis of CD11b-expressing cells in biological samples. It is designed for flow cytometry applications.
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Lab products found in correlation
Interferon γ apc xmg1.2, by Thermo Fisher Scientific (1 mentions)
Cd8α apc cy7, by Thermo Fisher Scientific (1 mentions)
Cd8α efluor450, by Thermo Fisher Scientific (1 mentions)
Foxp3 fitc fjk 16s, by Thermo Fisher Scientific (1 mentions)
Facscanto 2, by BD (1 mentions)
Aqua live dead dye, by Thermo Fisher Scientific (1 mentions)
Violet proliferation dye 450, by BD (1 mentions)
Cd3 fitc 17a2, by Thermo Fisher Scientific (1 mentions)
Protein labelling kit, by Thermo Fisher Scientific (1 mentions)
Lsrii fortessa x 20 flow cytometer, by BD (1 mentions)
Ghost violet 510 viability dye, by Cytek Biosciences (1 mentions)
Sa bv421, by BioLegend (1 mentions)
2 protocols using cd11b percpcy5.5 m1 70
1
Multiparametric Flow Cytometry for Immune Cell Profiling
2
Multicolor Flow Cytometry Immunophenotyping
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Purified monoclonal antibody was directly conjugated to Alexa Fluor-488 by protein labelling kit (ThermoFisher Scientific) according to the manufacturer's protocol. Splenocytes were isolated from NOD mice, red blood cells lysed and resuspended at in complete DMEM at 6 × 106 cells ml−1. p63 or OVA141–160 was added to the media at a final concentration of 40 μM and the cells were incubated for 1.5 h at 37 °C with 5% CO2. Cells were collected incubated in 2.4G2 for 10 min on ice and stained at 1:100 dilution for 30 min at 4 °C with FS1-AF488 and antibodies against CD4-BUV395 (GK1.5, BD Biosciences), CD8α-APC-ef780 (53–6.7, eBioscience), CD3ɛ-BV650 (145-2C11, BD Biosciences), B220-PE (RA3-6B, Tonbo), CD11c-PE-Cy (N418, eBioscience), CD11b-PerCp-Cy5.5 (M1/70, Tonbo), F4/80-APC (BM8.1, Tonbo), IAg7-biotin (10.2-16, BioXcell), SA-BV421 (BioLegend) and dead cells were gated out using Ghost violet 510 viability dye (Tonbo), and run on a LSRII Fortessa X-20 flow cytometer (Becton Dickinson) and analyzed using FlowJo software (v10).
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