Whole blood stainings were performed within 24 h after blood collection. Leukocytes were counted using Cell-Dyn Sapphire Hematology System (Abbott Diagnostics, Chicago, IL, USA). For staining, 1 million cells per tube were used based on direct blood cell count. Directly labeled antibodies were added to whole blood and incubated for 20 min at 4 °C, followed by 10 min red-blood-cells lysis (Bühlmann Laboratories, Schönenbuch, Switzerland) and subsequently washed using cold PBS. All anti-human antibodies were used at the concentrations recommended by the manufacturer: anti-CD15-PeCy7 (clone HI98), anti-CD14-Pe (clone MφP9), anti-CD163-FITC (clone GHI/61), anti-CD11b-BV510 (clone ICRF44), anti-CD33-V450 (clone WM53), anti-CD64-APCH7 (clone 10.1), anti-CD117-APC (clone YB5.B8), anti-CD45RA-PeCy7 (clone HI100), anti-CD25-Pe (clone M-A251), anti-CD4-FITC (clone RPA-T4), anti-CD8-V500 (clone SK1), anti-CD45RO-BV421 (clone UCHL1), anti-CD3-APCH7 (clone SK7), and CD127-Alexa Fluor 647 (clone HIL-7R-M21) and 7AAD (all from Becton Dickinson). BD FACSCanto II (Becton Dickinson) instrument was used to analyze samples and FlowJo 10.6.2 (Treestar Inc., Ashland, OR, USA) software and several software plugins (FlowCLEAN, downsample_V3, FlowSOM, tSNE) were used to analyze all data.
Anti cd11b bv510
Manufactured by BD
Sourced in United States
Anti-CD11b-BV510 is a fluorochrome-conjugated antibody that binds to the CD11b cell surface antigen. CD11b is expressed on the surface of various immune cells, including monocytes, macrophages, and neutrophils. The BV510 fluorochrome allows for the detection and analysis of CD11b-positive cells using flow cytometry.
Automatically generated - may contain errors
Lab products found in correlation
Cell dyn sapphire hematology system, by Abbott (2 mentions)
Anti cd15 pecy7, by BD (2 mentions)
Anti cd45ro bv421, by BD (2 mentions)
Facscanto 2, by BD (2 mentions)
Anti cd3 apc h7, by BD (2 mentions)
Anti cd45ra pe cy7, by BD (2 mentions)
Anti cd25 pe, by BD (2 mentions)
Anti cd4 fitc, by BD (2 mentions)
Anti cd8 v500, by BD (2 mentions)
Anti cd117 apc, by BD (2 mentions)
Anti cd163 fitc, by BD (2 mentions)
Lsrfortessa flow cytometer, by BD (2 mentions)
Anti cd45 v500, by BD (2 mentions)
Anti ly6g bv605, by BD (2 mentions)
Pe anti f4 80, by BD (2 mentions)
Lsrfortessa, by BD (2 mentions)
Anti cd16 32, by BD (2 mentions)
Cytofix cytoperm kit, by BD (2 mentions)
7 aad, by BD (1 mentions)
Anti cd14 pe, by BD (1 mentions)
6 protocols using anti cd11b bv510
1
Comprehensive Leukocyte Immunophenotyping Protocol
2
Whole Blood Immune Cell Profiling
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Whole blood staining's were performed within 24 hours after blood collection. Leukocytes were counted using Cell-Dyn Sapphire Hematology System (Abbott Diagnostics, Chicago, IL, USA). For staining, 1 million cells per tube were used. Directly labelled antibodies were added to whole blood and incubated for 20 minutes at 4 °C, followed by 10 minutes red-blood-cells lysis (Bühlmann Laboratories, Schönenbuch, Switzerland) and washing using cold PBS. All anti-human antibodies were used at the concentrations recommended by the manufacturer: anti-CD15-PeCy7 (clone HI98), anti-CD14-Pe (clone MφP9), anti-CD163-FITC (clone GHI/61), anti-CD11b-BV510 (clone ICRF44), anti-CD33-V450 (clone WM53), anti-CD64-APCH7 (clone 10.1), anti-CD117-APC (clone YB5.B8), anti-CD45RA-PeCy7 (clone HI100), anti-CD25-Pe (clone M-A251), anti-CD4-FITC (clone RPA-T4), anti-CD8-V500 (clone SK1), anti-CD45RO-BV421 (clone UCHL1), anti-CD3-APCH7 (clone SK7), and CD127-Alexa Fluor 647 (clone HIL-7R-M21) and 7AAD (all from Becton Dickinson). BD FACSCanto II (Becton Dickinson) instrument was used to analyse samples and FlowJo 10.6.2 (Treestar Inc., Ashland, OR, USA) software and several software plugins (FlowCLEAN, downsample_V3, FlowSOM, tSNE) were used to analyse all data.
3
Peripheral Blood Leukocyte Profiling by Flow Cytometry
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Blood samples were collected by cardiac puncture into EDTA-coated probes. Then, 50 µL of blood sample was incubated with 25 µL of blocking buffer (20% rat serum and 20% FBS in PBS) for 10 min at room temperature (RT). After incubation, surface antigens were stained with the following antibodies: anti-CD11b BV510 (562950; BD Horizon; 1:400), anti-CD45 V500 (562129; BD Horizon; 1:200), anti-Ly6G BV605 (563005; BD Horizon; 1:200) and Ly6C BV711 (128037; BioLegend (San Diego, CA, USA); 1:400) for 30 min at room temperature (RT) in the dark. Subsequently, 500 µL of RBC Easy Lyse 1× solution (S2364; Dako (Glostrup, Denmark)) was added to the samples to lyse erythrocytes, and the samples were lysed for 15 min at 4 °C. After passing the cells through a 100 μm strainer, the percentages of peripheral blood leukocytes subpopulations were analysed with a BD LSRFortessa flow cytometer.
4
Phenotypic Characterization of Murine Splenocytes and Neutrophils
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Splenocytes or isolated bone marrow neutrophils were seeded into 96-well plates (1–2 × 106 per well). Cells were centrifuged for 5 min at 350 RCF; then, they were stained with Fixable Viability Stain 440UV (565388; BD Horizon) according to the manufacture’s recommendation and incubated for 6 min at 37 °C, 5% CO2. After two washes, the samples were blocked with 5% FBS in PBS for 10 min at RT. Subsequently, surface antigens were labelled using the following antibodies: anti-CD11b BV510 (562950; BD Horizon; 1:400), anti-CD45 V500 (562129; BD Horizon; 1:200), anti-Ly6G BV605 (563005; BD Horizon; 1:200), anti-Ly6C BV711 (128037; BioLegend; 1:400), anti-CD80 Pe-Cy7 (104734; BioLegend; 1:200), anti-CD83 APC (558206; BD Pharmingen; 1:200), anti-I-A/I-E BB700 (746197; BD OptiBuild; 1:200) and anti-CD86 FITC (561962; BD Pharmingen; 1:200) for 30 min at RT. Cells were then fixed using BD Cell fix and analysed using a BD LSRFortessa flow cytometer.
5
Lung Single Cell Preparation and Intracellular Staining
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Lung single cells were prepared as previously described (Longhi et al., 2009 (link)). Briefly, lungs were digested in RPMI 1640 supplemented with 20μg/L Liberase (Roche) and 25μg/L DNase I (Roche) at 37°C for 30min. After red blood cells were removed using the ammonium-chloride-potassium buffer, lung single cells were blocked using anti-CD16/32 (553142, BD Biosciences) and stained with BV510-anti-CD11b (562950, BD Biosciences), APC-anti-CD11c (17-0114-81, eBiosciences), PE-anti-F4/80 (565410, BD Biosciences), BV650-anti-Ly6G (740554, BD Biosciences), and PE-Cy7-anti-Ly6C (4341610, Invitrogen). The cells were subsequently fixed and permeablized using BD CytoFix/CytoPerm Kit (BD Biosciences) according to the manufacturer’s instructions. Intracellular virus staining was performed using an anti-IAV M2 antibody (Abcam) and a FITC conjugation kit (Abcam). Flow cytometry was performed using BD LSRFortessa (BD Biosciences) and raw data were analyzed using FlowJo software.
6
Multiparametric Immune Cell Analysis
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Splenocytes and lung cells were incubated with anti-CD16/32 (BD Bioscience, USA) for 30min at 4°C in PBS supplemented with 2% FBS (FACS buffer) to block non-specific binding with Fc receptors. Cells were surface-stained with BV650-anti-CD8 (BD-563234, USA), BV711-anti-CD4 (BD-563050, USA), BV510anti-CD3 (BD-563024, USA), PECY7-anti-CD25 (eBioscience-25-0251-82, USA), PE-anti-F4/80 (BD-565410, USA), BV510-anti-CD11b (BD-562950, USA), APC-anti-CD11c (eBioscience-17-0114-81, USA) for 20 min on ice and washed twice with PBS. For intracellular staining, cells were fixed and permeablized using BD CytoFix/CytoPerm Kit (BD Bioscience, USA) and stained with APC-anti-IL-4 (eBioscience-17-7041-82, USA), PE-anti-IFN-γ (eBioscience-12-7311-82, USA), PE-anti-IL-2 (eBioscience-12-7021-82, USA), BV421-anti-IL-17 (BD-563354, USA), PECY7-anti-TNF-ɑ (BD-557644, USA), Pacific blue-anti-Foxp3 (eBioscience-48-5773-82, USA) for 20min. Cells were washed with 1×BD Perm/Wash buffer and were finally resuspended in 4% paraformaldehyde (PFA) for flow cytometry analysis. Flow cytometry was performed using BD LSR Fortessa (BD Bioscience, USA) and raw data was analyzed using FlowJo software.
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