Cd115 bv421

Manufactured by BioLegend

The CD115 BV421 is a fluorescently-labeled antibody that binds to the CD115 surface antigen. CD115 is also known as the macrophage colony-stimulating factor receptor (M-CSFR) or CSF-1R. This antibody can be used for the identification and analysis of cells expressing CD115 by flow cytometry.

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4 protocols using cd115 bv421

Multiparameter Flow Cytometry of Hematopoietic Cells

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Single cell suspensions obtained from colony forming assays or hematopoietic organs from mice were surface stained with monoclonal antibodies. Antibodies used for analysis of human immature stem and progenitor cells from colony forming assays: CD34 PE-Cy7(581), CD38 APC (HIT2), CD10 PE/PeCy5/APC (HI10a), CD45RA PerCP-Cy5.5 (HI100), CD90 APC-Cy7 (5E10) (Biolegend). Antibodies for erythroid cells from colony assays and HEMA: CD117 PE-Cy7 (104D2), CD71 APC/APC-Cy7 (CY1G4) (Biolegend), CD235a BV421 (HIR2) (BD biosciences). Antibodies for myeloid cells from colony forming assays: CD34 PE-Cy7 (581), CD33 PE/PerCP-Cy5.5 (WM53), CD14 APC (M5E2), CD115 BV421 (g-4D2-1E4), CD15 PeCy5 (W6D3), CD66b PerCP-Cy5.5 (G10F5) (Biolegend). Antibodies for megakaryocyte populations: CD41 PerCP-Cy5.5 (HIP8), CD61 APC (VI-PL2) (Biolegend). Antibodies for overall human and human myeloid cells in xenograft assay: CD45 Biotin (HI30), CD34 PE-Cy7/APC-Cy7 (581), CD33 PE/PerCP-Cy5.5 (WM53), CD15 PeCy5 (W6D3). Antibodies for human B cells in xenograft assay: CD19 PE-Cy7 (HIB19), IgM APC-Cy7 (MHM-88), CD10 APC (HI10a) (Biolegend). Antibody to detect murine cells in xenograft assay: CD45 PE-Cy7/PerCP-Cy5.5 (30-F11) (Biolegend). Streptavidin APC/V450 (Biolegend) were used as secondary antibodies. BD LSRFortessa was used for flow cytometry. Analyses were performed using FlowJo.

Afreen S., Bohler S., Müller A., Demmerath E.M., Weiss J.M., Jutzi J.S., Schachtrup K., Kunze M, & Erlacher M. (2020). BCL-XL expression is essential for human erythropoiesis and engraftment of hematopoietic stem cells. Cell Death & Disease, 11(1), 8.

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Omentum Immune Cell Characterization

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Mice were sacrificed and whole omenta were excised. Samples were placed in 6ml polypropylene tubes and blocked using FC Block (PharMingen, San Diego, CA) at 10μg/ml in 200 μl of FACs buffer and placed on a shaker for 15 minutes at 4°C. Following incubation, samples were stained with the following antibodies for 2 hours at 4°C: CD11b-PE (eBioscience; Clone M1/70), CD102-Alexa Fluor 647 (BioLegend; Clone 3C4), and CD115-BV421 (BioLegend; Clone AFS98) or left unstained. Samples were washed twice by the addition of 4ml FACs buffer and rotated at 4°C for 30 minutes. After the final wash, samples were collected from the tubes using a snipped off P1000 tip and placed onto glass slides with FACs buffer. A cover slip was placed on top of the omentum and pressed down. Samples were viewed via fluorescence microscopy and digital images were acquired.

Martin A.T., Giri S., Safronova A., Eliseeva S.I., Kwok S.F, & Yarovinsky F. (2024). Parasite-induced IFN-γ regulates host defense via CD115 and mTOR-dependent mechanism of tissue-resident macrophage death. PLOS Pathogens, 20(2), e1011502.

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Multiparametric Flow Cytometry of Murine Hematopoietic Cells

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Single cell suspensions obtained from colony-forming assays or hematopoietic organs from mice were surface stained with monoclonal antibodies: CD34 PE-Cy7(581), CD38 APC (HIT2), CD10 PE/APC (HI10a), CD45RA PerCP-Cy5.5 (HI100), CD90 APC-Cy7 (5E10), CD117 PE-Cy7 (104D2), CD71 APC (CY1G4), CD33 PE (WM53), CD14 APC (M5E2), CD115 BV421 (4D2-1E4), CD15 PeCy5 (W6D3), CD66b PerCP-Cy5.5 (G10F5), CD19 PE-Cy7 (HIB19), IgM APC-Cy7 (MHM-88), CD45 Biotin (HI30), CD45 PE-Cy7/V500 (30-F11) (Biolegend), and CD235a BV421 (HIR2) (BD Biosciences). Streptavidin PerCP-Cy5.5/V450 (Biolegend) was used as a secondary antibody. BD LSRFortessa and FlowJo were used for flow cytometry and analyses, respectively. The gating strategy was published earlier.^{31 (link)}

Bohler S., Afreen S., Fernandez-Orth J., Demmerath E.M., Molnar C., Wu Y., Weiss J.M., Mittapalli V.R., Konstantinidis L., Schmal H., Kunze M, & Erlacher M. (2020). Inhibition of the anti-apoptotic protein MCL-1 severely suppresses human hematopoiesis. Haematologica, 106(12), 3136-3148.

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Isolation and Characterization of Peritoneal Immune Cells

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Peritoneal exudate cells (PECs) were isolated via lavage and single cell suspensions were briefly washed with PBS prior to red blood cell lysis using ACK lysis buffer. Prepared cells were stained with Zombie Yellow (BioLegend) to assess live status of cells. The following antibodies were used, though not necessarily all in the same panel: F4/80- BV785 (BioLegend; Clone BM8), CD115-BV421 (BioLegend; Clone AFS98), CD11c-BV421 (BD Bioscience; Clone HL3), CD115-BUV395 (BD Bioscience; Clone AFS98), CD19-BUV737 (BD Bioscience; Clone 1D3), CD226-PE (BD Bioscience; Clone TX42.1), CD11b-PE (eBioscience; Clone M1/70), MHCII (I-A/I-E)- PE/Cy7 (BioLegend; Clone M5/114.15.2), Ly6G-PE/CF594 (BD Bioscience; Clone 1A8), Ly6C-APC/eFluor780 (eBiosceince; Clone HK1.4), CD102-Alexa Fluor 647 (BioLegend; Clone 3C4 (mlC2/4)), CD102-Alexa Fluor 488 (eBioscience; Clone 3C4 [mlC2/4]), CCR2-FITC (BioLegend; Clone SA203G11), and CD11b-PErCP/Cy5.5 (BioLegend; Clone M1/70). Antibody cocktails were prepared in FACs staining buffer (phosphate-buffered saline, 1% FBS, 1% 0.5mM EDTA). Cellular fluorescence was measured using an LSRII Fortessa flow cytometer, and data were analyzed using FlowJo software (Tree Star).

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