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Anti cd11b clone m1 70

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Anti-CD11b (clone M1/70) is a monoclonal antibody that specifically binds to the CD11b antigen expressed on the surface of certain immune cells, such as macrophages and neutrophils. It can be used for the detection and analysis of these cell populations in various research applications.

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Lab products found in correlation

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Spelling variants of this product

CD11b (454 citations) Anti-CD11b (175 citations) CD11b (M1/70, (58 citations) CD11b (clone M1/70, (43 citations) M1/70 (41 citations) Anti-CD11b (M1/70, (34 citations) Clone M1/70 (19 citations) V450 anti-CD11b (clone M1/70; (2 citations) Rat monoclonal anti-CD11b (Clone M1/70, (2 citations) APC-Cy7 rat anti-CD11b (clone M1/70, (2 citations)

35 protocols using anti cd11b clone m1 70

1

Enriching Immune Cell Populations from Tumor Samples

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CD11b+ splenocytes and CD45+ population from TC-1 tumors were enriched using positive selection by labeling cells with biotin conjugated anti-CD11b (clone M1/70, BD Biosciences, Carlsbad, CA) and anti-biotin Miltenyi magnetic beads (130-090-485, Miltenyi Biotec, GE) or CD45 Microbeads (130-052-301, Milteny Biotec, GE), respectively. Post sort analysis was performed to evaluate final enrichment of the target population. In general, CD45+ cells displayed 95% or more viability and 90% enrichment, and we could enrich splenocytes preparations to approximately 40% CD11b+ cells (S1 Fig) and over 95% viable cells. Tumor associated macrophages comprised approximately 85% of the total leukocyte infiltrate in the tumors, CD45+ (a representation of these populations in the total tumor suspension can be seen in S1 Fig). The CD45- population includes tumor cells, endothelial cells and fibroblasts.
Silveira C.R., Cipelli M., Manzine C., Rabelo-Santos S.H., Zeferino L.C., Rodríguez Rodríguez G., de Assis J.B., Hebster S., Bernadinelli I., Laginha F., Boccardo E., Villa L.L., Termini L, & Lepique A.P. (2019). Swainsonine, an alpha-mannosidase inhibitor, may worsen cervical cancer progression through the increase in myeloid derived suppressor cells population. PLoS ONE, 14(3), e0213184.
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2

Isolation of Murine Stomach Immune Cells

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Whole mouse stomachs were harvested and processed using Miltenyi’s Gentle Dissociator (Miltenyi Biotec, Boston, MA). In brief, the stomach was cut into 5 mm pieces and then transferred to a C-tube (Miltenyi Biotec) in 5 ml RPMI/10% FBS. The preset Miltenyi Biotec program m_imptumor 02 was run once and then the tissue was digested for 30 min at 37°C in a solution containing (0.32 mg/ml Dispase and 0.30 mg/ml Collagenase D, Roche) while shaking in a CO2 incubator. After the 37°C incubation, 100U/ml of DNase (Sigma) was added to each tube and the Miltenyi Gentle Dissociator was run for a second and third time using the preset program m_imptumor 02. The tissue homogenate was passed through a 70 μm cell strainer (BD). Cells were harvested by centrifugation, washed, and then live cells were counted by using a hemocytometer and trypan blue exclusion staining. The samples were stained with 2 μg/ml anti- F4/80, 1.5 μg/ml anti-Gr1 (clone RB6-8C5), and 2 μg/ml anti-CD11b (clone M1/70) (all antibodies were purchased from BD Biosciences, and analyzed on a BD LSR II flow cytometer).
Beceiro S., Radin J.N., Chatuvedi R., Piazuelo M.B., Horvarth DJ J.r., Cortado H., Gu Y., Dixon B., Gu C., Lange I., Koomoa D.L., Wilson K.T., Scott Algood H.M, & Partida-Sánchez S. (2016). TRPM2 Ion Channels Regulate Macrophage Polarization and Gastric Inflammation During Helicobacter pylori Infection. Mucosal immunology, 10(2), 493-507.
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3

Brain Tissue Dissociation and Cell Isolation

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Blood was prepared by ACK lysis. Single cell suspensions of brain were prepared as previously described (17 (link), 38 (link)). Mice were euthanized with CO2 and transcardially perfused with ice-cold HBSS. Brains were grossly free of blood. Whole brain was minced, and either mechanically triturated (in experiments for determination of relative proportions of infiltrating cell types) or enzymatically homogenized (in cell sorting experiments) with Neural Tissue Dissociation Kit with Papain, Miltenyi) and passed through a 70 μm cell strainer to form a single cell suspension. Cells were enriched by centrifugation over a discontinuous gradient of HBSS, 37% and 70% Percoll and collecting the fraction at the 37%/70% boundary. Cells were then washed and stained with fluorophore conjugated antibodies prior to analysis on a FACSAria II flow cytometer and cell sorter (BD). Antibodies include anti-CD11b (clone M1/70, BD), anti-CD45 (clone 30-F11, BD), anti-CD64 (clone X54-5/7.1, BD), anti-Ly6G (clone 1A8, Biolegend), anti-Ly6C (clone HK1.4, Biolegend). Propidium iodide was used as a live/dead marker. Gating was performed as described in Figure S1.
Denstaedt S.J., Spencer-Segal J.L., Newstead M.W., Laborc K., Zhao A.P., Hjelmaas A., Zeng X., Akil H., Standiford T.J, & Singer B.H. (2018). S100A8/A9 drives neuroinflammatory priming and protects against anxiety-like behavior after sepsis. Journal of immunology (Baltimore, Md. : 1950), 200(9), 3188-3200.
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4

Isolation and Analysis of Pancreatic Macrophages

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Pancreatic cells were isolated by collagenase digestion and macrophages were identified by surface staining with anti-F4/80 (clone BM8, eBioscience) and anti-CD11b (clone M1/70, BD Biosciences) (30 (link)). Intracellular staining was performed with anti-TNF-α (clone MP6-XT22, R&D Biosystems) and the appropriate isotype control (R&D Biosystems). Samples were collected (400,000 events) on an Attune NxT flow cytometer (Thermo Scientific) and analyzed with FlowJo version 10.0.7 Software (Treestar Incorporated) following the gating scheme (Suppl. Fig. 1).
Burg A.R., Das S., Padgett L.E., Koenig Z.E, & Tse H.M. (2017). Superoxide Production by NADPH Oxidase Intensifies Macrophage Anti-Viral Responses during Diabetogenic Coxsackievirus Infection. Journal of immunology (Baltimore, Md. : 1950), 200(1), 61-70.
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5

Isolation and characterization of brain immune cells

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The brain tissue was removed at necropsy after intravascular perfusion. For isolation of cells from the brain, the brain was carefully freed of meninges. Brain immune cells were isolated from all areas of the brain, by enzymatic digestion of minced tissue, followed by Percoll (Sigma-Aldrich) gradient, as described previously [27 (link)]. The isolated cells were quantified in a Z2 Coulter Counter (Beckman Coulter, Brea, CA), and further characterized by flow cytometry, using anti-CD11b (clone M1/70, BD Pharmingen, San Diego, CA), anti-CD45LCA (clone 2B11, BD Biosciences, San Diego, CA), anti-CD14-PE (clone M5E2, BD Pharmingen, San Diego, CA), anti-CD16-FITC (clone 3G8, BD Pharmingen), anti-monkey CD3-biotin (clone FN-18, Invitrogen Biosource, Carlsbad, CA) followed by streptavidin-PerCP or -APC (BD Pharmingen), anti-human CD8-PE, −FITC, or -PeCy5 (clone DK25, Dako, Carpinteria, CA), anti-CCR5, CCR2, CX3CR1, CD80 (BD Biosciences, San Jose, CA), and CD44v6 (clone 2 F10, Zymed, San Francisco, CA).or isotype controls (BD Pharmingen). Stained cells were acquired by a FACSCalibur (BD Biosciences, San Jose, CA) flow cytometer, and analyzed in FlowJo 6.2.1 software (Tree Star Inc., Ashland, OR), as previously described [27 (link)].
Najera J.A., Bustamante E.A., Bortell N., Morsey B., Fox H.S., Ravasi T, & Marcondes M.C. (2016). Methamphetamine abuse affects gene expression in brain-derived microglia of SIV-infected macaques to enhance inflammation and promote virus targets. BMC Immunology, 17, 7.
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6

Immune Cell Profiling from Brain Tissue

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Brains were isolated and homogenized, and single-cell suspensions were prepared by filtration through a 100 μm mesh. Cellular infiltrates were stained with anti-CD45.2 (clone 104, BD Biosciences), anti-CD11b (clone M1/70, BD Biosciences), anti-Ly-6G/Ly-6C (clone RB6-8C5, BD Biosciences), and/or TLR2 (clone 6C2, eBioscience), and acquired on a FACS Canto (BD Biosciences) running DIVA software.
Balathasan L., Tang V.A., Yadollahi B., Brun J., Labelle M., Lefebvre C., Swift S.L, & Stojdl D.F. (2017). Activating Peripheral Innate Immunity Enables Safe and Effective Oncolytic Virotherapy in the Brain. Molecular Therapy Oncolytics, 7, 45-56.
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7

Isolation of Murine Stomach Immune Cells

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Whole mouse stomachs were harvested and processed using Miltenyi’s Gentle Dissociator (Miltenyi Biotec, Boston, MA). In brief, the stomach was cut into 5 mm pieces and then transferred to a C-tube (Miltenyi Biotec) in 5 ml RPMI/10% FBS. The preset Miltenyi Biotec program m_imptumor 02 was run once and then the tissue was digested for 30 min at 37°C in a solution containing (0.32 mg/ml Dispase and 0.30 mg/ml Collagenase D, Roche) while shaking in a CO2 incubator. After the 37°C incubation, 100U/ml of DNase (Sigma) was added to each tube and the Miltenyi Gentle Dissociator was run for a second and third time using the preset program m_imptumor 02. The tissue homogenate was passed through a 70 μm cell strainer (BD). Cells were harvested by centrifugation, washed, and then live cells were counted by using a hemocytometer and trypan blue exclusion staining. The samples were stained with 2 μg/ml anti- F4/80, 1.5 μg/ml anti-Gr1 (clone RB6-8C5), and 2 μg/ml anti-CD11b (clone M1/70) (all antibodies were purchased from BD Biosciences, and analyzed on a BD LSR II flow cytometer).
Beceiro S., Radin J.N., Chatuvedi R., Piazuelo M.B., Horvarth DJ J.r., Cortado H., Gu Y., Dixon B., Gu C., Lange I., Koomoa D.L., Wilson K.T., Scott Algood H.M, & Partida-Sánchez S. (2016). TRPM2 Ion Channels Regulate Macrophage Polarization and Gastric Inflammation During Helicobacter pylori Infection. Mucosal immunology, 10(2), 493-507.
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8

Single-cell isolation from brain and spleen

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Brain single cell suspensions were prepared as previously described (17 ). Perfused spleens were dissected, minced and gently macerated through 40 μm cell strainer to form a single cell suspension. Strainers were washed with 30 mL of PBS with 1% bovine serum albumin, 2 mM EDTA, 25 mM HEPES. Cells were washed, Fc receptors blocked, and stained with fluorophore conjugated antibodies prior to analysis on a FACSAria II flow cytometer and cell sorter (BD). Antibodies included anti-CD11b (clone M1/70, BD), anti-CD45 (clone 30-F11, BD), anti-Ly6G (clone 1A8, Biolegend), anti-Ly6C (clone HK1.4, Biolegend). Gating strategy for myeloid cells in the brain and spleen available in supplemental data (Figure S1).
Denstaedt S.J., Spencer-Segal J.L., Newstead M., Laborc K., Zeng X., Standiford T.J, & Singer B.H. (2020). Persistent neuroinflammation and brain specific immune priming in a novel survival model of murine pneumosepsis. Shock (Augusta, Ga.), 54(1), 78-86.
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9

Characterizing Lung Immune Cell Types

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Single-cell lung suspensions were prepared as previously described 34 (link). Innate immune cells were classified based on surface expression of the following markers: AMs: CD11chiCD11b−, PMNs: Ly6GhiCD11bhi, LMs: CD11c−Ly6G−F4/80+CD11b+, iMs: CD11c−Ly6G−F4/80+CD11b+Ly6Chi. The following mAbs were used for these investigations: anti-Ly6G (clone 1A8; BD), anti-Ly6C (clone AL-21; BD), anti-CD11b (clone M1/70; BD), anti-F4/80 (clone BM8; ebiosciences), and anti-CD11c (clone HL3; ATCC).
For intracellular cytokine staining (ICS), 2–4 × 106 lung cells were cultured for 6 h in medium containing Brefeldin A, either alone or supplemented with purified protein derivative (SSI). Cells were phenotyped according to their CD4 and CD8 surface expression (anti-CD4, clone RM4–5; BD), (anti-CD8, clone 53–6.7, BD) and staining intracellularly with anti-IFN-γ (clone XMG1.2, BD) and anti-TNF-α (clone XT-22, ATCC) Abs. Stained cells were acquired using a BD FACS Canto II instrument. Data analysis was carried out using BD FACSDiva and FACS Analyzer software.
Dorhoi A., Yeremeev V., Nouailles G., Weiner J., Jörg S., Heinemann E., Oberbeck-Müller D., Knaul J.K., Vogelzang A., Reece S.T., Hahnke K., Mollenkopf H.J., Brinkmann V, & Kaufmann S.H. (2014). Type I IFN signaling triggers immunopathology in tuberculosis-susceptible mice by modulating lung phagocyte dynamics. European Journal of Immunology, 44(8), 2380-2393.
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10

Multiparametric Flow Cytometry Analysis

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Flow cytometry analysis was performed by Attune Acoustic Focusing Cytometer (Life Technologies) using FlowJo software (Tree Star). For Intracellular cytokine staining, cells were stimulated with 20 ng/mL phorbol 12‐myristate 13‐acetate (Sigma) and 1 mmol/L ionomycin (Sigma) for 5 hour in the presence of a GolgiStop (BD Bioscience). The antibodies used were as follows; anti‐CD16/CD32 (clone 2.4G2; BD Bioscience), anti‐CD4 (clone H129.19; BD Bioscience), anti‐CD25 (clone PC61; BD Bioscience), anti‐CD103 (clone M290; BD Bioscience), anti‐GITR (clone DTA1; BD Bioscience), anti‐CTLA‐4 (clone UC10; BD Bioscience), anti‐Foxp3 (clone FJK‐16s; eBioscience), anti‐CD11c (clone HL3; BD Bioscience), anti‐CD80 (clone 16‐10A1; BD Bioscience), anti‐CD86 (clone GL1; BD Bioscience), anti‐CD49b (clone HMa2; BD Bioscience), anti‐LAG3 (clone C9B7W; BD Bioscience), anti‐CD11b (clone M1/70; BD Bioscience), anti‐Ly6C (clone AL‐21; BD Bioscience), anti‐CD115 (clone AFS98; eBioscience), anti‐F4/80 (clone BM8; eBioscience), anti‐CD206 (clone C068C2; BioLegend), anti‐IFNγ (clone XMG1.2; eBioscience), anti‐IL‐4 (clone BVD4‐1D11; eBioscience), anti‐IL‐10 (clone JES5‐16E3; eBioscience) and isotype‐matched control antibodies.
Kasahara K., Sasaki N., Yamashita T., Kita T., Yodoi K., Sasaki Y., Takeda M, & Hirata K. (2014). CD3 Antibody and IL‐2 Complex Combination Therapy Inhibits Atherosclerosis by Augmenting a Regulatory Immune Response. Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease, 3(2), e000719.
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