Anti cd11b apc clone m1 70

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Anti-CD11b-APC (clone M1/70) is a fluorescently-labeled monoclonal antibody that binds to the CD11b cell surface antigen. CD11b is a component of the Mac-1 integrin complex and is expressed on the surface of myeloid cells, including monocytes, macrophages, and granulocytes. The APC (allophycocyanin) fluorescent label allows for detection of CD11b-positive cells using flow cytometry.

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CD11b (M1/70, (82 citations) Anti-CD11b-APC (46 citations) CD11b (clone M1/70, (44 citations) M1/70 (35 citations) Anti-CD11b (clone M1/70, (25 citations) Anti-CD11b (M1/70) (24 citations) Clone M1/70 (15 citations) CD11b-APC (M1/70, (9 citations) CD11b APC (clone M1/70), (8 citations) Anti-CD11b-APC (M1/70, (2 citations)

5 protocols using anti cd11b apc clone m1 70

Flow Cytometric Immunophenotyping of Macrophages

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Flow cytometry was performed as previous reported methods.^{38 (link)–40 (link)} In brief, single-cell suspensions from fresh mouse CLNs or cultured BM-macrophages were filtered by a 70-μm cell strainer, centrifuged, and resuspended. Cells were incubated with a stain kit (LIVE/DEAD Fixable Yellow Dead Cell Stain Kit, L34959, Thermo Fisher Scientific, Waltham, MA), followed by staining with anti-CD11b-APC (clone M1/70; eBioscience, San Diego, CA), anti-F4/80-APC/Cy7 (clone BM8; Biolegend), anti-TSLPR-PE (FAB5461P; R&D Systems, Minneapolis, MN), and anti-OX40L-PE/Cy7 (clone RM134L; Biolegend).
The gating strategy used in this study was as follows: dead cells were excluded by gating live dye negative cells, subsequently gated on the basis of forward scatter height versus forward scatter area (singlets 1), then gated on side scatter height versus side scatter area (singlets 2). Cells were then gated on F4/80⁺CD11b⁺ cells, and frequencies of F4/80⁺TSLPR⁺ and F4/80⁺ OX40L⁺ were recorded. Negative controls consisted of FMO cells. A BD Canto II Benchtop cytometer (Becton Dickinson, San Jose, CA) was used for flow cytometry, and data were analyzed with BD Diva software v6.7 (BD Biosciences, San Jose, CA) and FlowJo software v10. (Tree Star Inc., Ashland, OR).

Deng R., Chen X., Zhang Y., Bian F., Gao N., Hu J., Wang C., de Souza R.G., Lu F., Pflugfelder S.C, & Li D.Q. (2019). Short ragweed pollen promotes M2 macrophage polarization via TSLP/TSLPR/OX40L signaling in allergic inflammation. Mucosal immunology, 12(5), 1141-1149.

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Lung Single Cell Isolation and Characterization

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Preparation of lung single cells was described by Dr Smiley (16 (link)). In brief, cells isolated from lung tissues were ground with glass rod and digested with collagenase, and then single cells were incubated with Fc Block (clone 2.4G2) for 15 min at 4°C, washed 3 times. For enumeration of CD45 cells, macrophages, and neutrophils, pulmonary lymphocytes were stained on ice with anti-CD45-EF450 (cloneRM-5), anti-CD11b-APC (clone M1/70), and anti-Ly6G-FITC (clone 1A8), anti-Gr-1-Alexafluor700 (RB6-8C5), anti-CD3-PerCP-eFlour™-710 (clone17A2), anti-CD4-eFlour^®450 (cloneRM4-5), anti-CD8a-PE-Cy7 (clone53-6.7) (eBioscience, USA). Data were gated for forward scatter/side scatter and collected on an FACSCanto II (BD Biosciences) and analyzed using FlowJo software (Tree Star).

Sun Y., He Z., Li J., Gong S., Yuan S., Li T., Ning N., Xing L., Zhang L., Chen F., Li Z., Wang J., Luo D, & Wang H. (2021). Gentamicin Induced Microbiome Adaptations Associate With Increased BCAA Levels and Enhance Severity of Influenza Infection. Frontiers in Immunology, 11, 608895.

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Infiltrating Brain Immune Cell Analysis

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Six days post-infection, mice were exsanguinated and perfused as above; brains were harvested and homogenized in RPMI media-containing 0.5 mg/mL collagenase (Gibco LifeTechnologies), 0.01 mg/mL DNAse I (Roche) and 2 mM EDTA. Infiltrating cells were separated on a 33.3 % Percoll solution. Cells were stained for flow cytometry analyses with the following; Zombie Aqua Dye-V500 (BioLegend), anti-CD45-APC-eFluor780 (clone 30-F11, eBioscience), anti-CD4-PerCP Cyanine5.5 (clone RM4-5, eBioscience), anti-CD8alpha-PE (clone 53-6.7, eBioscience), anti-TCRbeta-FITC (clone H57-597, eBioscience), anti-CD19-BV421 (clone 6D5, BioLegend), F4/80-eFluor450 (clone BM8, eBioscience), anti-CD11b-APC (clone M1/70, eBioscience), anti-CD11c-PerCP Cyanine5.5 (clone N418, eBioscience), anti-Ly6C-PE (clone HK1.4, eBioscience), and anti-Ly6G-FITC (clone 1A8, BioLegend). Leukocytes were gated as CD45^hi cells.

Moradin N., Torre S., Gauthier S., Tam M., Hawari J., Vandercruyssen K., De Spiegeleer B., Fortin A., Stevenson M.M, & Gros P. (2016). Cysteamine broadly improves the anti-plasmodial activity of artemisinins against murine blood stage and cerebral malaria. Malaria Journal, 15, 260.

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Multicolor Flow Cytometry for Cell Identification

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Cell identification was performed according to a previous study with slight modifications [29] (link). In brief, cells were kept on ice, and all incubations were performed at 4 °C. Fc receptors were first blocked with anti-CD16/32 antibodies (clone 93, eBioscience). Cells (1 × 10⁶) were then incubated with the following fluorophore-labeled antibodies specific for cell-surface components: anti-F4/80-FITC (clone BM8, eBioscience) and anti-CD11b-APC (clone M1/70, eBioscience) in flow buffer (1% bovine serum albumin and 0.09% sodium azide in PBS) for 30 min. After that, cells were washed with PBS and fixed with 4% paraformaldehyde in PBS for 30 min. Finally, cells were washed and resuspended in PBS and stored at 4 °C until analysis by flow cytometry.

Wu Y., Gong X., Shen J, & Zhu K. (2023). Postantibiotic leukocyte enhancement-mediated reduction of intracellular bacteria by macrophages. Journal of Advanced Research, 58, 117-128.

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Comprehensive Immune Cell Profiling

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Flow cytometry analysis of splenic and peritoneal B cell subsets was performed as described previously 6 (link), 13 (link) using directly conjugated antibodies on single cell suspensions of freshly isolated spleens and peritoneal cells. Peripheral blood was diluted with PBS + 2% dextran (Sigma) for at least 30 minutes at 37°C to concentrate the RBCs at the bottom of the tube. The upper clear phase was collected, and cells were incubated with blocking anti-CD16/32 antibody (clone 93; eBiosciences). Peripheral monocytes and neutrophils were identified by staining with anti-CD11b APC (clone M1/70; eBiosciences), anti-Ly6C FITC (clone HK1.4; Biolegend) and anti-Ly6G-PE (clone 1A8; Biolegend). Resident peritoneal and splenic macrophages were stained with an anti-F4/80 antibody conjugated to APC (clone BM8; Biolegend). To quantify TACI expression, cells were stained with anti-TACI PE / APC and isotype control (anti-mouse IgD; clone 11-26c; eBiosciences) where indicated. T regulatory cells were quantified using the Treg detection kit (Miltenyi). Data were acquired on a FACS Calibur (BD) or LSRII Fortessa (BD) and were analyzed using Flow Jo software 7.6 (Treestar).

Tsiantoulas D., Sage A.P., Göderle L., Ozsvar-Kozma M., Murphy D., Porsch F., Pasterkamp G., Menche J., Schneider P., Mallat Z, & Binder C.J. (2018). B Cell–Activating Factor Neutralization Aggravates Atherosclerosis. Circulation, 138(20), 2263-2273.

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