Anti human cd11b pe

Manufactured by Thermo Fisher Scientific

Sourced in United Kingdom, United States

The anti-human CD11b-PE is a fluorescently-labeled antibody that specifically binds to the CD11b antigen expressed on the surface of certain immune cells, such as monocytes and macrophages. It can be used in flow cytometry applications to identify and quantify these cell populations.

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6 protocols using anti human cd11b pe

Delineating Monocyte Subsets with Fluorescent Probes

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Fluorescent immune cell probes for the delineation of monocyte subsets included anti-human CD14-V450, anti-human CD16-FITC, anti-human HLA-DR-PerCP-Cy5.5, anti-human CD33-APC, anti-human CD11b-PE, and IgG isotype controls (ebioscience or BD Biosciences). Cells were counted using an LSRII flow cytometer with the data was analyzed with BD FACSDIVA software. The submitted flow cytometry gating paradigm (Additional file 2: Figure S2) depicts cell populations for analysis of mature monocyte populations and the immature MDSC population.

Thome A.D., Faridar A., Beers D.R., Thonhoff J.R., Zhao W., Wen S., Pascual B., Masdeu J.C, & Appel S.H. (2018). Functional alterations of myeloid cells during the course of Alzheimer’s disease. Molecular Neurodegeneration, 13, 61.

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Flow Cytometry Analysis of Immune Cells

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Flow cytometry analyses were performed using either an LSR Model II BD FACSArray (BD Biosciences, Oxford, UK) or a Novocyte (Acea Biosciences, San Diego, CA, USA) flow cytometer. Cell-sorting experiments were performed using either Influx or FACSAria fluorescence-activated cell sorters (both from BD Biosciences). Antibodies used were anti-human CD11b-PE, anti-human CD14-FITC (fluorescein isothiocyanate), anti-human CD86-PerCP-eFluor710, and anti-Human CD117-PE (eBioscience, Hatfield, UK).

Maiques-Diaz A., Spencer G.J., Lynch J.T., Ciceri F., Williams E.L., Amaral F.M., Wiseman D.H., Harris W.J., Li Y., Sahoo S., Hitchin J.R., Mould D.P., Fairweather E.E., Waszkowycz B., Jordan A.M., Smith D.L, & Somervaille T.C. (2018). Enhancer Activation by Pharmacologic Displacement of LSD1 from GFI1 Induces Differentiation in Acute Myeloid Leukemia. Cell Reports, 22(13), 3641-3659.

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Comprehensive Flow Cytometry Staining Protocol

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All flow cytometry data were acquired using either using LSR II SORP or LSR Fortessa flow cytometers (BD Biosciences). All staining was carried out in FACS buffer (2% FBS in PBS) for 30 min on ice unless otherwise described. The following antibodies were used anti-human CD235a-APC (eBioscience, Clone HIR2), anti-human CD71-FITC (eBioscience, Clone OKT9), anti-human CD71-PEcy7 (eBioscience, Clone OKT9), ant-human CD49d-PE (Miltenyi, Clone MZ18-24A9), anti-human CD41a-PE (eBioscience, Clone HIP8), anti-human CD11b-PE (eBioscience, Clone ICRF44), anti-mouse Ter119-APC (eBioscience, Clone TER119), anti-mouse CD71-PE (eBioscience, Clone R17217) and Alexa Fluor-647 anti-phospho STAT5 (pY694) (BD Bioscience Cat#: 612599). Hoechst 33342 (Life Technologies, H1399) was used to visualize nuclei.

Nandakumar S.K., McFarland S.K., Mateyka L.M., Lareau C.A., Ulirsch J.C., Ludwig L.S., Agarwal G., Engreitz J.M., Przychodzen B., McConkey M., Cowley G.S., Doench J.G., Maciejewski J.P., Ebert B.L., Root D.E, & Sankaran V.G. (2019). Gene-centric functional dissection of human genetic variation uncovers regulators of hematopoiesis. eLife, 8, e44080.

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METTL3 Knockout in CD11b Cells

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Cas9 expressing cells were transduced with gRNA vectors targeting the catalytic domain of METTL3 or empty vectors and stained with anti-mouse CD11b PE/Cy5 (Biolegend, cat. no. 101210) and anti-human CD11b PE (eBiosciences, cat. no. 9012-0118). Data were analysed by using LSRFortessa (BD) and FlowJo.

Barbieri I., Tzelepis K., Pandolfini L., Shi J., Millán-Zambrano G., Robson S.C., Aspris D., Migliori V., Bannister A.J., Han N., De Braekeleer E., Ponstingl H., Hendrick A., Vakoc C.R., Vassiliou G.S, & Kouzarides T. (2017). Promoter-bound METTL3 maintains myeloid leukaemia via m6A-dependent translation control. Nature, 552(7683), 126-131.

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Ortho-topolin riboside modulates STAT3 signaling

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Ortho-topolin riboside (oTR, purity >99%) was obtained from OlChemim GmbH (Czech Republic). RPMI 1640 and fetal bovine serum (FBS) were obtained from GIBCO (USA). Anti-STAT3, anti-phospho-STAT3^Y705, anti-JAK2, anti-phospho-JAK2^Y1007/1008, anti-phospho-SHP-1^Tyr564, anti-phospho-SHP-2^Tyr542, anti-SHP-1, anti-SHP-2, and β-actin were obtained from Cell Signaling (USA). Wright-Giemsa staining solution was obtained from Sigma-Aldrich Corporation (USA). Penicillin-streptomycin, Cell Counting Kit-8 (CCK-8), and phosphatase inhibitor complex were obtained from the Beyotime Institute of Biotechnology (Beijing, China). Anti-human CD11b-PE was obtained from eBioscience (USA).

Wang L., Cheng J., Lin F., Liu S., Pan H., Li M., Li S., Li N, & Li W. (2019). Ortho-Topolin Riboside Induced Differentiation through Inhibition of STAT3 Signaling in Acute Myeloid Leukemia HL-60 Cells. Turkish Journal of Hematology, 36(3), 162-168.

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AML Cell Apoptosis and Cycle Analysis

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Cells were transduced with gRNA vectors or treated with SPHINX31 and stained at the indicated time points with anti-mouse CD11b PE/Cy5 (Biolegend, cat. no. 101210) and anti-human CD11b PE (eBiosciences, cat. no. 9012-0118) or anti-human CD13 FITC (eBioscience, cat. no. 11-0138-42). Data were analyzed by using LSRFortessa (BD) and FlowJo.
Apoptosis levels were measured in human and/or mouse AML cells transduced with dual gRNA vectors (against SRPK1 and 3’ BCL2 enhancer) and/or treated with 1 or 3 μM SPHINX31 (Exonate) at indicated time points, by using Annexin V (Life Technologies, cat. no. V13242). Data were analyzed by using LSRFortessa (BD) instruments.
Cell cycle stages were measured in human and/or mouse AML cells transduced with dual gRNA vectors against SRPK1 and/or treated with 1 or 3 μM SPHINX31 (Exonate) at indicated time points, using Propidium Iodide from Abcam (ab14083). Data were analyzed using LSRFortessa (BD) instruments.

Tzelepis K., De Braekeleer E., Aspris D., Barbieri I., Vijayabaskar M.S., Liu W.H., Gozdecka M., Metzakopian E., Toop H.D., Dudek M., Robson S.C., Hermida-Prado F., Yang Y.H., Babaei-Jadidi R., Garyfallos D.A., Ponstingl H., Dias J.M., Gallipoli P., Seiler M., Buonamici S., Vick B., Bannister A.J., Rad R., Prinjha R.K., Marioni J.C., Huntly B., Batson J., Morris J.C., Pina C., Bradley A., Jeremias I., Bates D.O., Yusa K., Kouzarides T, & Vassiliou G.S. (2018). SRPK1 maintains acute myeloid leukemia through effects on isoform usage of epigenetic regulators including BRD4. Nature Communications, 9, 5378.

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