Fitc anti human cd11b

Manufactured by BioLegend

FITC anti-human CD11b is a fluorescently labeled antibody that specifically binds to the CD11b cell surface antigen expressed on various immune cells, including monocytes, macrophages, and neutrophils. It is a useful tool for the identification and enumeration of these cell populations in flow cytometry applications.

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CD11b (457 citations) Anti-CD11b (179 citations) CD11b-FITC (96 citations) Anti-CD11b-FITC (57 citations) FITC anti-mouse/human CD11b antibody (9 citations) FITC anti-mouse/human CD11b (6 citations) FITC anti-human CD11b antibody (3 citations) FITC-conjugated anti-human CD11b (3 citations) FITC)-conjugated anti-mouse/human CD11b (2 citations) FITC-conjugated anti-human antibody directed against CD11b/Mac-1 (2 citations)

5 protocols using fitc anti human cd11b

BMSC-Derived Macrophage Phenotyping via FACS

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The expression of CD11b, CD206, CD11c, and F4/80 in BMSC-derived macrophages were analyzed using the FACS Calibur (Becton Dickinson, United States) and the CellQuest software (Becton Dickinson). The adherent cells were washed with PBS and collected using Accutase (Nacalai Tesque, Kyoto, Japan) and resuspended in 50 μL staining buffer (BD Pharmingen, Franklin Lakes, NJ, United States). The harvested cells were blocked with 2 μL Human Trustain FcX (Fc receptor Blocking Solution; BioLegend) for 10 min at room temperature, and stained with 2 μL antibodies in the dark for 30 min at 4°C. FITC anti- mouse CD11b, FITC anti-human CD11b, Alexa Fluor^®488 anti-human CD11b, BV421 anti-mouse CD206, PE anti-mouse F4/80 and APC anti-mouse CD11c (BioLegend).

Chen G., Sun Q., Cai Q, & Zhou H. (2022). Outer Membrane Vesicles From Fusobacterium nucleatum Switch M0-Like Macrophages Toward the M1 Phenotype to Destroy Periodontal Tissues in Mice. Frontiers in Microbiology, 13, 815638.

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Flow Cytometric Characterization of hADSCs

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The hADSCs of passages 3 or 5 were collected with 1×Tryple Express and centrifuged at 400 g for 5 min. After washing twice with 1×DPBS, cells were resuspended and incubated with pre-labelled antibodies for 15 min at room temperature. After two washes with 1×PBS, cells were resuspended in 300 μL 1×PBS and analyzed using a flow cytometer (BD FACSCalibur). Histograms were generated using the CELLQuest Pro software (BD Biosciences). The antibodies used were as follows: FITC Mouse IgG1,k,Iso-type Ctrl (FC) (BioLegend, 400110), FITC anti-human CD34 (BioLegend, 343504), FITC anti-human CD45, (BioLegend, 304006), FITC anti-human CD11b (BioLegend, 301330), FITC anti-human HLA-DR (BioLegend, 307604), FITC anti-human CD73 (BioLegend, 344016), FITC anti-human CD90 (BioLegend, 328108), APC Mouse IgG1,k,Isotype Ctrl (BioLegend, 400120), APC anti-human CD19 (BioLegend, 363006), PE Mouse IgG1,k,Isotype Ctrl (FC) (BioLegend, 400114), PE anti-CD105 (Endoglin) (BioLegend, 800504).

Yan K., Zhang J., Yin W., Harding J.N., Ma F., Wu D., Deng H., Han P., Li R., Peng H., Song X, & Kang Y.J. (2022). Transcriptomic heterogeneity of cultured ADSCs corresponds to embolic risk in the host. iScience, 25(8), 104822.

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Immunostaining of Tight Junction Proteins

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hAELVi and differentiated THP-1 cells were washed with PBS, fixed with 3.6 % para formaldehyde (PFA) (Sigma-Aldrich, 47608) in PBS for 15 min at RT, following a rinse with PBS. Next, the cells were permeabilized with 0.5% Triton X-100 (Sigma-Aldrich, T8787) in PBS for 5 min at RT. The cells were washed with PBS and blocked with 10% serum in PBS for 1 hour. For DAPI nucleic acid staining cells were incubated with DAPI solution (ThermoFisher scientific, D1306), diluted with PBS (ratio of 1:1000) for 5 minutes at RT. For tight junction proteins, Zonula occludens-1 (ZO-1) and Occludin staining, cells were incubated with the primary antibodies rabbit anti-ZO1 (ThermoFisher scientific, 617300)/mouse anti-Occludin (ThermoFisher scientific, 331500) diluted with PBS (ratio of 1:200) overnight at 4°C, followed by incubation with secondary antibody Alexa Fluor 488 anti-rabbit/ anti-mouse (Jackson ImmunoResearch, 111-545-144/ 115-545-062), diluted with PBS (ratio of 1:400) for 1 hour at RT. The differentiation of THP-1 towards macrophage-like cells was examined with FITC anti-human CD11b (BioLegend, 301329), diluted with PBS (ratio of 1:20) for 1 hour at RT. Confocal Microscopy imaging of fluorescent immunostaining was performed (Nikon Eclipse Ti with spinning disk, Yokogawa Japan).

Artzy-Schnirman A., Zidan H., Elias-Kirma S., Ben-Porat L., Tenenbaum-Katan J., Carius P., Fishler R., Schneider-Daum N., Lehr C.M, & Sznitman J. (2019). Capturing the onset of Bacterial Pulmonary Infection in Acini-on-Chips. Advanced biosystems, 3(9), e1900026.

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Polarization of THP-1 Monocytes to M1 and M2 Macrophages

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THP-1, the most widely used model for the human monocytes/macrophages, was obtained from Cell Bank of Chinese Academy of Sciences (Shanghai, China), and routinely cultured in RPMI 1640 medium (HyClone, USA) supplemented with 10% FBS and 1% penicillin/streptomycin. We polarized THP-1 monocytes according to the procedure reported previously [25 (link)]. THP-1 were firstly differentiated into M0 macrophages with 100 ng/ml phorbol 12-myristate 13-acetate (PAM, Sigma, USA) for 24 h. M0 cells were then polarized into M1 macrophages with incubation with 100 ng/ml LPS (PeproTech, USA) and 20 ng/ml IFN-γ (PeproTech) for an additional 48 h. To generate M2 phenotype, M0 populations were exposed to 20 ng/ml IL-4 (PeproTech) and 20 ng/ml IL-13 (PeproTech) for another 48 h. The phenotypes of polarized macrophages were determined by cell makers via flow cytometry. M1 and M2 macrophages were stained using lineage-specific antibodies, with PE anti-human CD86 for M1, APC anti-human CD206 (BioLegend) for M2, and FITC anti-human CD11b (BioLegend) for all monocytes. FlowJo software was used for statistical analysis.

Shen M., Yu H., Jin Y., Mo J., Sui J., Qian X, & Chen T. (2022). Metformin Facilitates Osteoblastic Differentiation and M2 Macrophage Polarization by PI3K/AKT/mTOR Pathway in Human Umbilical Cord Mesenchymal Stem Cells. Stem Cells International, 2022, 9498876.

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Macrophage Polarization Analysis

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Macrophage polarization After the cell pellet was collected and resuspended, FITC anti-human CD11b (301329, Biolegend) and APC anti-human CD86 (305411, Biolegend) were added for 30 min of incubation at 4 °C in the dark. Subsequently, 500 μL of Fixation Buffer was added for 20 min of incubation at room temperature. After centrifugation, the pellet was washed, resuspended, and then incubated with Intracellular Staining Perm Wash Buffer. Following another round of centrifugation, the cells were resuspended again and added with PE anti-human CD206 (321105, Biolegend) for 30 min of incubation at 4 °C. After centrifugation, the cells were washed with Intracellular Staining Perm Wash Buffer, centrifuged again, and resuspended. Finally, the cells were analyzed using flow cytometry.

Zhao X., Li M., Lu Y., Wang M., Xiao J., Xie Q., He X, & Shuai S. (2024). Sirt1 inhibits macrophage polarization and inflammation in gouty arthritis by inhibiting the MAPK/NF-κB/AP-1 pathway and activating the Nrf2/HO-1 pathway. Inflammation research : official journal of the European Histamine Research Society ... [et al.], 73(7).

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