Cd11b pe clone m1 70

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CD11b PE (clone M1/70) is a fluorescently labeled antibody that binds to the CD11b protein expressed on the surface of certain immune cells. It can be used to identify and quantify these cell populations in flow cytometry applications.

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CD11b (457 citations) CD11b (M1/70, (86 citations) CD11b-PE (81 citations) CD11b (clone M1/70, (72 citations) M1/70 (46 citations) Clone M1/70 (26 citations) CD11b PE (M1/70, (7 citations) CD11b-PE/Dazzle594 (clone M1/70, (3 citations) PE-Cy7 anti-mouse CD11b (clone M1/70, (2 citations) Anti CD11b-PE (Clone M1/70, (2 citations)

6 protocols using cd11b pe clone m1 70

Multiparametric Immune Cell Analysis

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Cells were stained using the following fluorophore conjugated anti-mouse antibodies: WGA-488 (Invitrogen), Ly6G-FitC (clone 1A8, BioLegend), Ly6G-BV510 (clone 1A8, BioLegend), Ly6G-PE (clone 1A8, BioLegend), CD11c-BV605 (clone N418, BioLegend), F4/80-FitC (clone BM8, BioLegend), F4/80-PE (clone BM8, BioLegend), CD11b-PE (clone M1/70, BioLegend), Ly6C-PE Cy7 (clone HK1.4, BioLegend), CD63-APC (clone NVG-2, BioLegend), CD9-APC (clone MZ3, BioLegend), IL-1α-PE (clone ALF-161, BioLegend), and IL-1β-APC (clone NJTEN3, ThermoFisher Scientific). Antibodies were diluted in wash buffer (PBS with 1% BSA and 2 mM EDTA). Cells were stained for 20 minutes at 4°C, washed with wash buffer, fixed for 15 minutes in BD Biosciences cytofix/cytoperm, and permeabilization prior to intracellular staining. ACEA Novocyte was used for flow cytometry, and Novoexpress software was used for subsequent analysis. AMNIS ImageStream was used for imaging flow cytometry and the AMNIS IDEAS software was used to calculate the colocalization coefficient.

Ratitong B., Marshall M, & Pearlman E. (2021). β-Glucan-stimulated neutrophil secretion of IL-1α is independent of GSDMD and mediated through extracellular vesicles. Cell reports, 35(7), 109139.

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Multiparametric Immune Cell Analysis

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Ratitong B., Marshall M, & Pearlman E. (2021). β-Glucan-stimulated neutrophil secretion of IL-1α is independent of GSDMD and mediated through extracellular vesicles. Cell reports, 35(7), 109139.

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Comprehensive Murine Immune Cell Profiling

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Blood samples collected from cardiac punctures were first processed by lysing red blood cells, followed by staining with the following antibodies: CD11c FITC (clone N418, TONBO #350114U100, 1:600), CD19 PerCP‐Cy5.5 (clone 1D3, TONBO #650193U100, 1:480), CD11b PE (clone M1/70, Biolegend #101207, 1:960), CD3e PE‐CD594 (clone 145‐2C11, BD Biosciences #562286, 1:120), CD62L PE‐Cy7 (clone MEL‐14, TONBO #600621U100, 1:600), CD4 APC (clone RM4‐5, Biolegend #100516, 1:480), CD8a A700 (clone 53‐6.7, Biolegend #100729, 1:600), Ly6G BV421 (clone 1A8, BD Biosciences #562737, 1:480), CD45 BV605 (clone 30‐F11, BD Biosciences #563053, 1:240), B220 BUV496 (clone RA3‐6B2, BD Biosciences #564662, 1:120). Samples were processed on a FACSymphony A5 cytometer and analyzed using FlowJo (v10) software.

Heuer S.E., Keezer K.J., Hewes A.A., Onos K.D., Graham K.C., Howell G.R, & Bloss E.B. (2023). Control of hippocampal synaptic plasticity by microglia–dendrite interactions depends on genetic context in mouse models of Alzheimer's disease. Alzheimer's & Dementia, 20(1), 601-614.

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Comprehensive Immune Cell Profiling in Lung Tissue

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Lung cells (1 × 10⁶ to 2 × 10⁶) were stained for surface antigens for 30 min at 4°C. Stained cells were washed and then fixed overnight in PBS containing 2% paraformaldehyde. The following antibodies were used: CD3-PE (clone 145-2C11; eBioscience), CD4-PB (clone RM4-5; eBioscience), CD8-FITC (clone 5H10-1; BioLegend), CD44-PerCPCy5.5 (clone 1M7; eBioscience), CD62L-PECy7 (clone MEL-14; BioLegend), CD19-APC (clone eBio1D3; eBioscience), Ly6G-APC (clone 1A8; BioLegend), Ly6C-PerCPCy5.5 (clone AL-21; BD Pharmingen), MHC II-FITC (clone AMS-32.1; BD Pharmingen), CD11b-PE (clone M1/70; BioLegend), and CD11c-PB (clone N418; BioLegend). Samples were run on an LSRII flow cytometer with Diva software, and the data were analyzed using FlowJo version 10.1.r7 software. The total number of cells in each gate was calculated using the total number of cells determined by the Countess automated cell counter. The gating strategy used is shown in Fig. S2 in the supplemental material.

Bhatt K., Machado H., Osório N.S., Sousa J., Cardoso F., Magalhães C., Chen B., Chen M., Kim J., Singh A., Ferreira C.M., Castro A.G., Torrado E., Jacobs WR J.r., Bhatt A, & Saraiva M. (2018). A Nonribosomal Peptide Synthase Gene Driving Virulence in Mycobacterium tuberculosis. mSphere, 3(5), e00352-18.

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Multicolor Flow Cytometry Analysis

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Blood collected from cardiac punctures were first processed by lysing red blood cells, followed by staining with the following antibodies: CD11c FITC (clone N418, TONBO, 1:600), CD19 PerCP-Cy5.5 (clone 1D3, TONBO, 1:480), CD11b PE (clone M1/70, Biolegend, 1:960), CD3e PE-CD594 (clone 145–2C11, BD Biosciences, 1:120), CD62L PE-Cy7 (clone MEL-14, TONBO, 1:600), CD4 APC (clone RM4–5, Biolegend, 1:480), CD8a A700 (clone 53–6.7, Biolegend, 1:600), Ly6G BV421 (clone 1A8, BD Biosciences, 1:480), CD45 BV605 (clone 30-F11, BD Biosciences, 1:240), B220 BUV496 (clone RA3–6B2, BD Biosciences, 1:120). Samples were processed on a FACSymphony A5 cytometer and analyzed using FlowJo (v10) software.

Heuer S.E., Keezer K.J., Hewes A.A., Onos K.D., Graham K.C., Howell G.R, & Bloss E.B. (2023). Genetic context controls early microglia-synaptic interactions in mouse models of Alzheimer’s disease. bioRxiv.

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Retina Immune Cell Isolation and Analysis

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Retinas were homogenized and pooled as previously described (Williams et al., 2017a (link)). Eyes were enucleated and placed directly into ice-cold HBSS. Retinas were dissected and placed into 100uL of HBSS with dispase (5U/mL) DNAse 1 (2000U/mL). Retinas were incubated for 20 minutes at 37°C whilst being shaken at 350RPM in an Eppendorf Thermomixer R. Samples were gently triturated. For flow cytometric analysis, each sample was stained with DAPI, CD45 BV605 (clone 30-F11, BD Biosciences, 1:240), and CD11b PE (clone M1/70, Biolegend, 1:960), followed by processing on a FACSymphony A5 cytometer, and analysis using FlowJo (v10) software.

Diemler C.A., MacLean M., Heuer S.E., Hewes A.A., Marola O.J., Libby R.T, & Howell G.R. (2024). Microglia Depletion leads to Increased Susceptibility to Ocular Hypertension-Dependent Glaucoma. bioRxiv.

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