Anti cd115 clone afs98

Manufactured by Thermo Fisher Scientific

Anti-CD115 (clone AFS98) is a laboratory reagent used for the identification and enumeration of cells expressing the CD115 antigen. It is designed for use in flow cytometry applications.

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3 protocols using anti cd115 clone afs98

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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Detailed FACS Sorting and Analysis

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For FACS sorting and analysis, we used previously described staining protocols (Mossadegh-Keller et al., 2013 (link)), published stem and progenitor cell definitions (Bryder et al., 2006 (link)), FACSCanto, LSRII, and FACSAriaIII equipment, and DIVA software (BD), analyzing only populations with at least 200 events. The following antibodies were used for staining cells: anti-CD117 (clone 2B8; BD), anti–Sca-1 (clone D7; BioLegend), anti-CD34 (clone RAM34; BD), anti-CD16/32 (clone 2.4G2; BD), anti-CD11b (clone M1/70; BD), anti-CD19 (clone 1D3; BD), anti-CD3e (clone 145-2C11; BioLegend), anti-Ly6G (clone 1A8; BioLegend), anti-Ly6C (clone HK1.4; BioLegend), anti-CD115 (clone AFS98; eBioscience), anti-CD45.2 (clone 104; BD), anti-CD45.1 (clone A20; BD), anti-B220 (clone RA3-6B2; eBioscience), anti-Ter119 (clone TER-119; eBioscience), and anti-CD71 (clone R17217; eBioscience). LIVE/DEAD Fixable Violet Dead cell dye (Invitrogen) was used as a viability marker. Information describing gating strategies is shown in Figs. S1 and S2.

Kandalla P.K., Sarrazin S., Molawi K., Berruyer C., Redelberger D., Favel A., Bordi C., de Bentzmann S, & Sieweke M.H. (2016). M-CSF improves protection against bacterial and fungal infections after hematopoietic stem/progenitor cell transplantation. The Journal of Experimental Medicine, 213(11), 2269-2279.

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Comprehensive Immune Cell Analysis by Flow Cytometry

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Whole blood was collected in an EDTA-buffered tube, subjected to red blood cell lysis, and centrifuged for 5 min at 300 × g, and cell pellets were subsequently stained with different antibody cocktails for analysis by flow cytometry. Flow cytometry was conducted with a FACS Canto II, using FACSDiva software (BD Biosciences). Cell populations were discriminated by the following antibody cocktail: anti-CD45 (clone 30-F11; eBioscience), anti-CD115 (clone AFS98; eBioscience), anti-Gr1 (clone RB6-8C5; Biolegend), anti-CD11b (clone M1/70; eBioscience), anti-B220 (clone RA3-6B2; eBioscience), and anti-CD3 (clone 145-2C11; eBioscience). Cell populations and marker expression were gated using the FlowJo analysis program (Treestar): leukocytes (CD45⁺), neutrophils (CD45⁺ CD115-Gr1^high), monocytes (CD45⁺ CD11b⁺ CD115⁺), and lymphocytes (CD45⁺ CD3⁺ and CD45⁺ B220⁺).

Kiouptsi K., Jäckel S., Pontarollo G., Grill A., Kuijpers M.J., Wilms E., Weber C., Sommer F., Nagy M., Neideck C., Jansen Y., Ascher S., Formes H., Karwot C., Bayer F., Kollar B., Subramaniam S., Molitor M., Wenzel P., Rosenstiel P., Todorov H., Gerber S., Walter U., Jurk K., Heemskerk J.W., van der Vorst E.P., Döring Y, & Reinhardt C. (2019). The Microbiota Promotes Arterial Thrombosis in Low-Density Lipoprotein Receptor-Deficient Mice. mBio, 10(5), e02298-19.

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