Cd11b apc m1 70

Manufactured by BD

CD11b-APC (M1/70) is a fluorescent-conjugated antibody that binds to the CD11b cell surface antigen. It is used in flow cytometry applications to identify and analyze cells expressing the CD11b receptor.

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4 protocols using cd11b apc m1 70

Multilineage Chimerism Analysis by Flow Cytometry

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Leukocytes were isolated from the blood, thymus, spleen, lymph node, BM, liver, and lung from mice that had received BM transplantation. After lysing red blood cells, the cells were washed and incubated for 20 min with Fc blocking antibody (Clone 93, 1:50 dilution, BioLegend, San Diego, CA) followed by the relevant antibody combinations (1:50 dilution) and FACS staining buffer for 30 min at 4 °C. Mixed chimerism was assessed in multiple lineages and compartments by staining cells with the following antibodies obtained from BioLegend (San Diego, CA) or BD Bioscience (San Jose, CA): H2-K^d FITC(SF1-1.1), CD3-PB (17A2), CD11b-APC (M1/70), CD19-APC-Cy7 (6D5), TER119-Alexa Fluor 700 (Ter-119), Gr-1-PE-Cy7 (RB6-8C5), CD49b-PE (DX5), SCA-1-APC-Cy7 (D7), Flt3/CD135-APC (A2F10), CD127/IL-7 alpha-Brilliant Violet 605 (A7R34), CD117/c-kit-APC (2B8), lineage cocktail-PB (17A2/RB6-8C5/RA3-6B2/Ter-119/M1/70), CD150-PE-Cy7 (TC15-12F12.2), and CD34-biotin (RAM34) plus streptavidin-PE. All data were collected using an LSRII flow cytometer (BD Biosciences) and analyzed with FlowJo^TM 10 software (version 10.2; Treestar, Ashland, OR) using previously published subset definitions^{14 (link)}.

Li Z., Czechowicz A., Scheck A., Rossi D.J, & Murphy P.M. (2019). Hematopoietic chimerism and donor-specific skin allograft tolerance after non-genotoxic CD117 antibody-drug-conjugate conditioning in MHC-mismatched allotransplantation. Nature Communications, 10, 616.

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Multi-Marker Immune Cell Profiling

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Tumor-infiltrating immune cells were incubated with Fc-blocking reagent (anti CD16/CD32, BD Biosciences), followed by CD11b-APC (M1/70, BD Biosciences), Gr1-FITC (RB6–8C5, BD Biosciences), CD11c-APC (HL3, BD Biosciences), MHC II-FITC (I-Ab, AF6–120.1, BD Biosciences), CD3 -APC (145–2C11, eBioscience), CD4-FITC (GK1.5, eBioscience), CD8a-APC (53–6.7, eBioscience) along with isotype-matched control. In vitro cultures dendritic cells (DCs) were stained with CD11b-APC, Gr1-FITC, CD11c-APC, MHC II-FITC, CD80-FITC (16–10A1, eBioscience) and CD86-FITC (GL1, eBioscience).

Foubert P., Kaneda M.M, & Varner J.A. (2017). PI3Kγ activates integrin α4 promotes immune suppressive myeloid cell polarization during tumor progression. Cancer immunology research, 5(11), 957-968.

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Identification of Mouse Hematopoietic Populations

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The following antibodies were used for identification of mature cell populations: CD41 PE (MWReg30, BD Biosciences), CD42d (1C2, Biolegend) CD11b APC (M1/70, BD Biosciences), Gr-1 PerCPCy5.5 (RB6-8C5, ebioscience), TER119 APC (TER119, BioLegend). For hematopoietic progenitor populations the following antibodies were used: APC lineage (Lin) markers (CD3, KT31.1; CD4, GK1.5; CD8, 53–6.7; B220, 6B2; Mac-1, M1/70; Gr-1, 8C5; and TER119, all from BD Biosciences), c-kit APC-Cy7 (2B8, BD Biosciences), CD34 PE (RAM34, BD Biosciences), CD16/32 PE-Cy7 (2.4G2, BD Biosciences), and Sca-1 Pacific Blue (D7, BioLegend). LKS, CMP, GMP, and MEP are defined as in^{15 (link)}. Cells were analyzed using an Aria III flow cytometer (BD Biosciences). Data was analyzed using FlowJo software (Treestar).

Brooks S.A., Luty S.B., Lai H.Y., Morse S.J., Nguyen T.K., Royer L.R., Agarwal A., Druker B.J, & Fleischman A.G. (2015). JAK2V617I results in cytokine hypersensitivity without causing an overt myeloproliferative disorder in a mouse transduction-transplantation model. Experimental hematology, 44(1), 24-9.e1.

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Infection and Flow Cytometry Analysis

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Mice were infected with 1×10³ CFUs flgB∷Tn10 pWSK129 (empty vector, kanamycin resistant) (WT) orflgB∷Tn10 pEM87 (FliC^ind plasmid, ampicillin resistant) (FliC^ind) S. Typhimurium in PBS by intraperitoneal (IP) injection, and treated with doxycycline as described above. Single cells were isolated from spleen, treated with Collagenase D, RBC lysis buffer, and flushed through a cell strainer (Falcon). Cells were Fc-blocked with anti-CD16/CD32 and stained with CD11b-APC (M1/70), Ly6G-APC Cy7 (1A8), NK-1.1-PE (PK136) (BD), and F4-80-PacBlue (BM8) (eBioscience), fixed and analyzed on a Becton Dickinson LRSIII (HTS) (UNC Flow Cytometry Core Facility).

Jorgensen I., Lopez J.P., Laufer S.A, & Miao E.A. (2016). IL-1β, IL-18, and eicosanoids promote neutrophil recruitment to pore-induced intracellular traps following pyroptosis. European journal of immunology, 46(12), 2761-2766.

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