Cd11b pb

Manufactured by Thermo Fisher Scientific

CD11b-PB is a fluorescently labeled monoclonal antibody that binds to the CD11b antigen, which is expressed on the surface of myeloid cells, including monocytes, macrophages, and granulocytes. The PB (Pacific Blue) fluorescent label allows for the detection and analysis of CD11b-positive cells using flow cytometry.

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2 protocols using cd11b pb

Flow Cytometry Analysis of Immune Cells

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Stained cells were analyzed using a BD LSR II-SORP system (Becton Dickinson) and analyzed with FlowJo software (Tree Star). The following mAbs were used: CD45-PerCPCy5, CD45-APC, CD8-APC, anti-IFNγ-PECy7 and anti-IL-4-FITC (BD Biosciences), Gr1-APC, CD11c-PECy7, CD11b-PB, F4/80-APC (eBiosciences), 1A8-APCCy7, Ly6C-FITC, 1A8-APCCy7, CD4-AF700. DAPI and AnV-FITC (BioLegends). In vivo bioluminescence imaging of MPO activity was quantified through injection of 200mg/kg luminol (Carbosynth) i.p. 10 minutes before luminescence acquisition. Photon emission was acquired for 10 minutes using a Xenogen IVIS Imaging system.

Hurrell B.P., Schuster S., Grün E., Coutaz M., Williams R.A., Held W., Malissen B., Malissen M., Yousefi S., Simon H.U., Müller A.J, & Tacchini-Cottier F. (2015). Rapid Sequestration of Leishmania mexicana by Neutrophils Contributes to the Development of Chronic Lesion. PLoS Pathogens, 11(5), e1004929.

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Multiparameter Flow Cytometry Analysis

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The antibodies used were as follows: CD19-PECy7 (1D3), CD8-APC (53-6.7), Ly6C-FITC (AL-21), CD68-PE (FA11), CD44-FITc (IM7), CD45-PerCP (30-F11), CD69-PE (H1.2F3), CXCR5-ef450 (SPRCL5), CD138-APC (281-2), CD21-PE (7G6), CD275-PE (HK5.3), CD122-APC (TM-b1), CD11b-PB (M1/70.15), IFNγ-eFluor 450 (XMG1.2), CD4-PerCP (L3T4), F4/80-APC-eFluor780 (BM8), Foxp3-PE (MF23), CD3-APC-CY7/eFluor780 (17A2), Ly-6G-PE (1A8), from eBioscience (San Diego, CA) and anti-mouse CD16/CD32 (The Lymphocyte Culture Centre, UVA, Charlottesville, VA). For some staining, we used: PerCP-CD4, biotinylated-CXCR5, followed by APC-streptavidin, or FITC-GL-7, PE-FAS and PerCP-B220 staining (all from Biolegend). To distinguish between live and dead cells, Viability Live Dead-ef650 (eBioscience, San Diego, CA) was used. Anti–CD3 and –CD28 were utilized for in vitro assays (eBioscience, San Diego, CA). Recombinant proteins used were as followed: mouse TGFβ and IL-2 was purchased from PeproTech (Rocky Hill, NJ). LPS was purchased through Sigma-Aldrich (St. Louis, MO).

Taghavie-Moghadam P.L., Wassem T.C., Hattler J., Glenn L.M., Dobrian A.D., Kaplan M.H., Yang Y., Nurieva R., Nadler J.L, & Galkina E.V. (2017). STAT4 regulates CD8+Treg/Tfh cell axis and promotes atherogenesis in insulin-resistant Ldlr−/− mice. Journal of immunology (Baltimore, Md. : 1950), 199(10), 3453-3465.

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