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Cd11b apc cy7

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CD11b-APC-Cy7 is a fluorochrome-conjugated antibody that binds to the CD11b cell surface antigen. CD11b is a component of the integrin Mac-1 (CD11b/CD18) and is expressed on the surface of various immune cells, including monocytes, macrophages, and neutrophils. The APC-Cy7 fluorochrome attached to the CD11b antibody can be used for multicolor flow cytometry applications.

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Lab products found in correlation

Cd45 vf450, by Cytek Biosciences (2 mentions) Neutrophil monocyte respiratory burst assay kit, by Cayman Chemical (1 mentions) Ghost dye 510, by Cytek Biosciences (1 mentions) Ly6c pe cf594, by BD (1 mentions) Cytoflex s flow cytometer, by Beckman Coulter (1 mentions) F4 80 apc cy7, by BioLegend (1 mentions) Zombie yellow fixable viability dye, by BioLegend (1 mentions) Cd138 bv711, by BD (1 mentions) Annexin 5 apc, by Thermo Fisher Scientific (1 mentions) Cd21 apc b ly4, by BD (1 mentions) Igm fitc, by Thermo Fisher Scientific (1 mentions) Cd45 2 percp cy5, by Cytek Biosciences (1 mentions) Cd43 pe, by BD (1 mentions) Cd90.2 apc cy7, by BioLegend (1 mentions) Fc block, by BD (1 mentions) Aria 2, by BD (1 mentions) Cd8 pecy7, by Cytek Biosciences (1 mentions) Ghost dye bv510, by Cytek Biosciences (1 mentions) Anti mouse cd16 cd32 fc blocked, by BD (1 mentions) Cd8 pe cy5, by Cytek Biosciences (1 mentions)

4 protocols using cd11b apc cy7

1

Profiling Immune Cell Functionality

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Single-cell brain and bone marrow suspensions were washed twice in 1X sterile, cold PBS, followed by viability staining with Ghost Dye 510 (Tonbo Biosciences) for 30 minutes in the dark at room temperature. Cells were centrifuged at 500g for 5 minutes at 4°C, the supernatant was discarded and the pelleted cells were re-suspended in 100ul of 1:100 FC Receptor Block (Tonbo Biosciences) for 10 minutes at room temperature. Cells were then stained with following surface antibodies: CD45-vf450, CD11b-APC-Cy7, Ly6G-Pe-Cy7, Ly6C-APC (Tonbo Biosciences) for 30 minutes at room temperature, protected from light. Following surface staining, samples were incubated with dihydrorhodamine 1,2,3 (DHR) and 200 nM PMA for 45 minutes at 37°C according to the manufacturer’s instructions (Neutrophil Monocyte Respiratory Burst Assay Kit, Cayman Chemical), then washed and analyzed immediately for rhodamine fluorescence on a Beckmann Coulter Cytoflex S Flow Cytometer. Data was analyzed using FlowJo (TreeStar).
Roy-O’Reilly M.A., Ahnstedt H., Spychala M.S., Munshi Y., Aronowski J., Sansing L.H, & McCullough L.D. (2020). Aging exacerbates neutrophil pathogenicity in ischemic stroke. Aging (Albany NY), 12(1), 436-461.
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2

Multiparametric Immune Cell Phenotyping

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Following this blocking step, cells were re-suspended in the following cocktail of fluorescently conjugated antibodies: CD45-vf450 (Tonbo Biosciences), CD11b APC-Cy7 (Tonbo Biosciences), Ly6G-PeCy (Tonbo Biosciences) and Ly6C-PeCF594 (BD Biosciences) and incubated for 30 minutes at room temperature. After staining, cells were washed twice and re-suspended in 300ul FACS buffer for analysis on a Cytoflex S Flow Cytometer (Beckmann Coulter). Data files were analyzed using FlowJo (TreeStar). The representative gating strategy used is given in Supplementary Figure 2. Positive and negative populations were defined using fluorescence minus one (FMO) controls.
Roy-O’Reilly M.A., Ahnstedt H., Spychala M.S., Munshi Y., Aronowski J., Sansing L.H, & McCullough L.D. (2020). Aging exacerbates neutrophil pathogenicity in ischemic stroke. Aging (Albany NY), 12(1), 436-461.
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3

Multicolor FACS Analysis of B Cell Subsets

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Cells were resuspended at a concentration of 106 cells/100 μl fluorescence-activated cell sorting (FACS) buffer (1X PBS, 2 mM EDTA, 1% BSA). Cells were stained with Fc block (BD Biosciences) for fifteen minutes, stained with antibody-fluorophores for one hour on ice, then washed with ten volumes of FACS buffer. The following antibodies were used: B220-PE-Cy7 (RA3–6B2, Tonbo), CD11b-APC-Cy7 (M1/70, Tonbo), CD45.1-FITC (A20, Tonbo), CD45.2-PerCP-Cy5.5 (104, Tonbo), CD90.2-APC-Cy7 (30-H12, BioLegend), F4/80-APC-Cy7 (BM8, BioLegend), CD138-BV711 (281–2, BD), CD21-APC (B-ly4, BD), CD43-PE (S7, BD), GL7-eFluor660 (GL-7, eBioscience), CD23-eFluor450 (B3B4, eBioscience), IgM-FITC (II/41, eBioscience), Annexin V-APC (kit number 88–8007-72, eBioscience), and Zombie Yellow Fixable Viability Dye (kit number 423104, BioLegend). An LSRII was used for analysis and an ARIAII was used for sorting (BD Biosciences). All flow cytometry data was analyzed with FlowJo version 9.9.5. Preceding all flow cytometry analyses presented is the following gating strategy: 1) lymphocytes (forward scatter [FSC]-area by side scatter [SSC]-area), 2) singlets (FSC-width by FSC-height), 3) singlets (SSC-width by SSC-height), 4) live cells (viability dye−), 5) exclusion of non-B cell lineage cells (Thy1.1−F4/80−CD11b−).
Haines R.R., Barwick B.G., Scharer C.D., Majumder P., Randall T.D, & Boss J.M. (2018). The histone demethylase LSD1 regulates B cell proliferation and plasmablast differentiation. Journal of immunology (Baltimore, Md. : 1950), 201(9), 2799-2811.
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4

Flow Cytometric Analysis of Lung Immune Cells

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The lungs were aseptically collected from mock and IAV-infected mice, and single cells were prepared after collagenase digestion of lungs, as previously described (PMC7336542). One million cells per sample were stained with Ghost Dye-BV510 (Tonbo Biosciences, San Diego, CA) and anti-mouse CD16/CD32 (FC blocked) (BD Biosciences, San Jose, CA) for 30 min at 4°C for 30 min. For cell surface staining, cells were incubated for 30 min at room temperature with anti-mouse CD3-APC-Cy7, CD3-AF488, CD4-BV786, CD8-PE-Cy7(Tonbo), CD8-PE-Cy5, CD44 PerP-Cy5.5, CD44 PE-Cy7, CD62L-FITC, CXCR3-BV-650, CXCR3-PE-Dazzle; For monocytes, cells were stained with Cd11b APC/Cy7, Ly6C BV711, Ly6G FITC and CCR2 PE/Cy7. All the antibodies were purchased from Biolegend unless specified.
Guo K., Yombo D.J.K., Xu J., Wang Z., Schmit T., Tripathi J.K., Hur J., Sun J., Olszewski M.A., & Khan M.N. (2022). The chemokine receptor CXCR3 promotes CD8+T cell-dependent lung pathology during influenza pathogenesis.
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