Cd11b

Manufactured by BD

Sourced in United States, Germany, Canada, Spain, France

CD11b is a cell surface receptor that is expressed on various immune cells, including monocytes, macrophages, and neutrophils. It is involved in cell-cell adhesion and migration processes.

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Lab products found in correlation

F4 80, by BD (71 mentions) Facscanto 2, by BD (26 mentions) Siglec f, by BD (25 mentions) Cd105, by BD (22 mentions) Facscalibur, by BD (21 mentions) F4 80, by Thermo Fisher Scientific (17 mentions) Mhcii, by BD (17 mentions) Sca 1, by BD (15 mentions) Hla dr, by BD (14 mentions) Facsdiva software, by BD (14 mentions) Ifn γ, by BD (14 mentions) Cd62l, by BD (14 mentions) Ter119, by BD (13 mentions) F4 80, by BioLegend (12 mentions) Cd206, by BioLegend (12 mentions) Cd206, by BD (12 mentions) Lsrfortessa, by BD (12 mentions) Facsaria, by BD (10 mentions) Il 17a, by BD (9 mentions) Facsaria 2, by BD (9 mentions)

Spelling variants of this product

CD11b-FITC (104 citations) Anti-CD11b-FITC (51 citations) Anti-CD11b-PE (48 citations) Anti-CD11b-APC-Cy7 (31 citations) CD11b PE-CF594 (7 citations) APC anti-mouse CD11b (6 citations) APC-conjugated anti-mouse CD11B (3 citations) PE-Cy7-conjugated anti-CD11b (clone M1/70); (3 citations) CD11b-fluorescein isothiocyanate (3 citations) CD45+CD11b (2 citations)

454 protocols using cd11b

Isolation and Characterization of Myeloid Cells

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Single cell suspensions were made from spleens or peripheral blood of normal and 4T1 tumor-bearing mice (13 (link)), as well as lung tissues (74 (link)). Cells were labeled with fluorescence-conjugated antibodies: Gr-1, CD11b, Ly6G, Ly6C, F4/80, AnnexinV, 7AAD (BD Pharmingen), and CCR1 (R&D system). For flow cytometry analysis, cells were run on a FACS Calibur or Fortessa flow cytometer (BD, San Jose, CA) and analyzed on FlowJo. For sorting, Gr-1+CD11b+ cells, CD11b+Ly6G+ cells, CD11b+Ly6C+ cells, and CD11b+F4/80+ cells were sorted from spleens of 4T1 tumor-bearing mice by FACSAria flow cytometer (BD) or MACS (Magnetic-activated cell sorting) according to manufacturer protocol (Miltenyi Biotec). For sorting human CD33+ myeloid cells, normal human whole blood was obtained from NIH blood bank in clinical center. Myeloid cells were enriched by Ficoll-Paque™ (GE Healthcare), then labeled with CD33 antibody and sorted with MACS (Miltenyi Biotec).

Yan H.H., Jiang J., Pang Y., Achyut B., Lizardo M., Liang X., Hunter K., Khanna C., Hollander C, & Yang L. (2015). CCL9 induced by TGF-β signaling in myeloid cells enhances tumor cell survival in the premetastatic organ. Cancer research, 75(24), 5283-5298.

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Epididymal Adipose Tissue SVC Isolation

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Epididymal visceral adipose tissue was mechanically chopped and then digested with collagenase Type 4, DNAse 1, and 0.5% fatty acid free bovine serum albumin in phosphate buffered saline (PBS) for 50 minutes at 37°C in an incubator-shaker (120 RPM). Next, 0.5M EDTA was added with RPMI media and the cells were passed through a 70-μm cell strainer rinsed with 2% fetal bovine serum (FBS) in PBS. The cells were centrifuged at 1400 RPM for 5 minutes at 4°C, primary adipocytes were aspirated off from the top layer of the supernatant and the pellet containing the stromal vascular cells (SVC) was then incubated with red blood cell lysis buffer for 1 minute on ice. Two percent FBS-PBS was added to the SVCs and centrifuged at 1400 RPM for 5 minutes at 4°C and then passed through a 40-μm cell strainer. SVCs were stained with the following fluorophore-tagged antibodies obtained from BioLegend (San Diego, CA): Zombie Aqua, CD45 (PerCP-Cy5.5), CD11b (FITC), CD19 (APC-Cy7), F4/80 (PE), and MHCII (BV421). The following SVC subsets were analyzed using a BD LSRII flow cytometer: CD45⁺CD11b⁻CD19⁺ (B cells), CD45⁺CD11b⁺F4/80⁺MHCII⁺ (macrophages), CD45⁺CD11b⁻F4/80⁺MHCII⁻ (macrophages). All data were analyzed in FlowJo and gates were drawn from fluorescence minus one controls.

Pal A., Al‐Shaer A.E., Guesdon W., Torres M.J., Armstrong M., Quinn K., Davis T., Reisdorph N., Neufer P.D., Spangenburg E.E., Carroll I., Bazinet R.P., Halade G.V., Clària J, & Shaikh S.R. (2020). Resolvin E1 derived from eicosapentaenoic acid prevents hyperinsulinemia and hyperglycemia in a host genetic manner. The FASEB Journal, 34(8), 10640-10656.

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Immunophenotyping of Bone Marrow Myeloid Cells

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Bone marrow myeloid cells were obtained by flushing tibiae and femurs with growth medium, and enriched for CD45 expression using CD45 MicroBeads (Miltenyi Biotec) and magnetic-activated cell sorting according to the manufacturer’s instructions. Cells were spun down and labelled with the following antibodies for 45 min at 4 °C: APC-CD11b (101211, BioLegend), FITC-Gr-1 (108405, BioLegend), FITC-F4/80 (123107, BioLegend), PE-CD115 (165203, BioLegend) and PerCP-CD19 (115531, BioLegend). The number of Gr-1⁺CD11b⁺ neutrophils, F4/80⁺CD11b⁺ macrophages, CD115⁺CD11b⁺ monocytes and CD19⁺ B cells was assessed by flow cytometry (BD FACSCanto, BD Biosciences) as described before^{49 (link)} and quantified using Kaluza software (Beckman Coulter).
Circulating myeloid cells in blood were analysed using the Sysmex XN1000 and XN2000 Hematology Analyzer (Sysmex Corporation; University Hospital Leuven).

Stegen S., Moermans K., Stockmans I., Thienpont B, & Carmeliet G. (2024). The serine synthesis pathway drives osteoclast differentiation through epigenetic regulation of NFATc1 expression. Nature Metabolism, 6(1), 141-152.

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Tumor Cell and Immune Cell Profiling

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As described previously35 , subcutaneous tumours with KP1.9 cells were harvested from C57BL/6 flanks 3 weeks after implantation, minced, and shaken at 600 r.p.m. with 0.2 mg ml⁻¹ collagenase type I (Worthington Biochemical Corporation) in RPMI-1640 for 30 min at 37 °C. Digested samples were filtered (70 μm BD Falcon strainer); washed in PBS with 0.5% BSA and 2 mM EDTA; incubated with Fc-block (TruStain fcX anti-mouse CD16/32; clone 93; Biolegend) for 15 min at 4 °C; and labelled with antibodies as indicated for 45 min at 4 °C. Flow cytometry (LSRII, BD Biosciences) labelled tumour cells (CD45⁻ EpCAM⁺), TAM (CD45⁺ CD11b⁺ Ly6C^- Lin^- CD11c⁺ F4/80⁺), lymphocyte-like cells (CD45⁺ CD11b^- Lin⁺), along with CD45- EpCAM- host-cell populations. Antibodies included EpCAM (clone G8.8; eBioscience); CD45 (clone 30-F11; Biolegend), F4/80 (clone BM8; Biolegend), CD11c (clone N418; Biolegend), Ly6C (clone HK1.4; Biolegend); and CD11b (clone M1/70; BD Biosciences). The lineage (Lin) antibody mix contained anti-CD90.2 (clone 53–2.1), anti-B220 (clone RA3-6B2), anti-NK1.1 (clone PK136), anti-CD49b (clone DX5), anti-Ter119 (cloneTER-119) and anti-Ly6G (clone 1A8) (all BD Biosciences). 7-aminoactinomycin D (7-AAD, Sigma Aldrich) excluded dead cells. VT680 fluorescence was directly assessed using the LSRII flow cytometer, FlowJo v.8.8.7 (Tree Star, Inc.) and MATLAB.

Cuccarese M.F., Dubach J.M., Pfirschke C., Engblom C., Garris C., Miller M.A., Pittet M.J, & Weissleder R. (2017). Heterogeneity of macrophage infiltration and therapeutic response in lung carcinoma revealed by 3D organ imaging. Nature Communications, 8, 14293.

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Circulating CD11b+ Cells in Glioma

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Flow cytometry analysis was performed as previously described (19). Peripheral blood drawn from human subjects with histologically-confirmed grade II (n=10)and IV (n=10) gliomas and healthy controls was centrifuged at 500 g for 10 min at 4°C. Isolation of white blood cells was performed using a Ficoll-Paque Gradient (GE Healthcare) and stained with CD11b⁺ antibody (BD Biosciences 1:50).Blood collected from non-tumor controls was used solely for the purpose of comparison between low- and high-grade counterparts for circulating CD11b⁺ cells. Blood and bone marrow from mice were harvested and stained with CD11b (BD Biosciences 1:200) and GR1 antibodies (BD Biosciences 1:100).

Rajappa P., Cobb W.S., Vartanian E., Huang Y., Daly L., Hoffman C., Zhang J., Shen B., Yanowitch R., Garg K., Cisse B., Haddock S., Huse J., Pisapia D.J., Chan T.A., Lyden D.C., Bromberg J.F, & Greenfield. J.P. (2016). Malignant astrocytic tumor progression potentiated by JAK-mediated recruitment of myeloid cells. Clinical cancer research : an official journal of the American Association for Cancer Research, 23(12), 3109-3119.

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Immunophenotyping of Immune Cells

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Single cell suspensions of spleen and LN were made in 1× phosphate-buffered saline (PBS; 0.137M NaCl, 2.7mM KCl, 5.3mM Na₂HPO₄, 1.8mM KH₂PO₄), 2% fetal bovine serum (FBS; Atlanta Biologicals, S11550), using 70-µm nylon cell strainers (Falcon, 352350). Peritoneal cells were isolated by lavage (1× PBS, 2% FBS). Cells were blocked for FCGR2B/CD32 (eBiosciences,14-0161-85) and stained with the following mouse Abs from eBiosciences: CD11c (48-0114-82), CD3 (48-0032-80), (eFluor 450), CD23 (11-0232-82), GR1/Ly6G (11-5931-82), (FITC), PDCA1 (46-3172-80), GR1/Ly6G (45-5931-80), (PerCP-Cy5.5), B220 (47-0452-82), CD11b (47-0112-82), (eFluor 780), and BD Biosciences: CD69 (551113) (PerCP-Cy5.5), CD80 (553769), CD11b (01715B), CD138 (553714), CD69 (553237), (PE), CD5 (55035), CD21 (558658), B220 (553092), (APC). Cells were analyzed using an LSRII, FACSDiva software (BD Biosciences). Secondary analysis was performed using FCS Express 4 Research Edition (De Novo Software).

Weindel C.G., Richey L.J., Mehta A.J., Shah M, & Huber B.T. (2016). Autophagy in dendritic cells and B cells is critical for the inflammatory state of TLR7-mediated autoimmunity. Journal of immunology (Baltimore, Md. : 1950), 198(3), 1081-1092.

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Isolation and Characterization of Activated Lung Macrophages

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Mice were sacrificed and lungs were aseptically removed. To prepare a single-cell suspension, lungs were minced and digested in 5% FBS/PBS solution containing 250 U/ml collagenase IV (Worthington Biochemical Corporation) and 20 U/ml DNase (Roche) for 60 min at 37°C. Lung tissues were then passed through a 70-µm cell strainer, and red blood cells were lysed with ammonium-chloride-potassium buffer.
For cell sorting, lung cell suspensions were blocked with anti–mouse CD16/32 followed by incubation with CD11b (BD) and CD80 (eBioscience) antibodies. CD11b⁺ mCherry⁺ CD80^high cells (activated population) and CD11b⁺ mCherry⁺ CD80^low cells (resting population) were sorted using a cell sorter (S3; Bio-Rad Laboratories) with a 100-µm nozzle. Cells were then surface stained with CD11b (BD), CD80 (eBioscience), CD40 (eBioscience), CD86 (eBioscience), and MHCII (eBioscience). For iNOS staining, surface-stained cells were fixed and permeabilized using an intracellular fixation and permeabilization buffer set (eBioscience). Cells were then incubated for 30 min with an iNOS antibody (eBioscience). Data were acquired using a flow cytometer (LSR II; BD) and analyzed with FlowJo software (Tree Star).

Liu Y., Tan S., Huang L., Abramovitch R.B., Rohde K.H., Zimmerman M.D., Chen C., Dartois V., VanderVen B.C, & Russell D.G. (2016). Immune activation of the host cell induces drug tolerance in Mycobacterium tuberculosis both in vitro and in vivo. The Journal of Experimental Medicine, 213(5), 809-825.

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Quantification of Immune Regulatory Cells

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For detection of myeloid derived suppressor cells (MDSCs), peripheral blood cells from DC vaccine-immunized and control mice were collected, and cells were stained for 30 min at 4°C with antibodies against specific cell markers, including FITC-conjugated anti-mouse CD11b (for cell surface), APC-Cy7 conjugated anti-mouse Ly-6C and PE conjugated anti-mouse Ly-6G, (both for intracellular staining). All three antibodies were obtained from Biolegend, (San Diego, CA). The percentages of monocytic and granulocytic MDSCs were gated on CD11b⁺Ly-6C⁺ cells and CD11b⁺ Ly-6G⁺ cells, respectively.[19 (link)] For Treg cell detection, cells were surface stained with FITC-conjugated anti-mouse CD25 and inter-cellularly stained with APC conjugated anti-mouse Foxp3, followed by cell permeabilization with the Cytofix/Cytoperm Plus kit according to the manufacturer’s protocol for all three tested antibodies, and the reagent kit from BD pharmingen (San Diego, CA). Percentage of Treg cells was gated on CD25⁺ Foxp3⁺ cells.[20 (link)] Fluorescence signals were detected by cytometry as described above.

Lin T.J., Liang W.M., Hsiao P.W., M. S P., Wei W.C., Lin H.T., Yin S.Y, & Yang N.S. (2015). Rapamycin Promotes Mouse 4T1 Tumor Metastasis that Can Be Reversed by a Dendritic Cell-Based Vaccine. PLoS ONE, 10(10), e0138335.

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Peritoneal Exudate Cell Analysis in Sepsis

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Peritoneal lavage was harvested before and after cecal slurry peritonitis-induced polymicrobial sepsis. Peritoneal exudate cells were incubated with FITC, PE, PerCP, or APC-conjugated anti-CD11b (BD Pharmingen, San Diego, CA), anti-CD11c (BioLegend, San Diego, CA), anti-F4/80 (BD Pharmingen), and anti-Gr1 (BioLegend) monoclonal antibodies. Subpopulations of PMNs (CD11b⁺F4/80⁻Gr1^hi) and macrophages (CD11b⁺F4/80⁺CD11c^lo) in the peritoneal cavity were analyzed by flow cytometry (BD Biosciences).

Zhou H., Lu X., Huang J., Jordan P., Ma S., Xu L., Hu F., Gui H., Zhao H., Bai Z., Redmond H.P., Wang J.H, & Wang J. (2022). Induction of Trained Immunity Protects Neonatal Mice Against Microbial Sepsis by Boosting Both the Inflammatory Response and Antimicrobial Activity. Journal of Inflammation Research, 15, 3829-3845.

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Tumor-associated Myeloid Subset Isolation

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Single cell suspensions were made from the lung tissues of tumor-bearing mice as described [50 (link)]. Gr-1+CD11b+, Ly6G+CD11b+Ly6C-F4/80-, Ly6C+CD11b+Ly6G-F4/80-, and F4/80+CD11b+ Ly6G-Ly6C- myeloid subsets were sorted from splenocytes, tumor tissues, and lungs as by FACSAria flow cytometer (BD). Antibodies including CD11b, Gr-1, Ly6G, Ly6C, F4/80, CD45, Lin, and CD31 were purchase from BD. CD117 was from eBioscience and Sca-1 was from Biolegend. FACS Calibur and Fortessa flow cytometer (BD, San Jose, CA) were used.

Jian J., Pang Y., Yan H.H., Min Y., Achyut B.R., Hollander M.C., Lin P.C., Liang X, & Yang L. (2016). Platelet factor 4 is produced by subsets of myeloid cells in premetastatic lung and inhibits tumor metastasis. Oncotarget, 8(17), 27725-27739.

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