Cd9 apc

Manufactured by BioLegend

Sourced in United States

CD9-APC is a fluorochrome-conjugated monoclonal antibody that binds to the CD9 antigen. CD9 is a member of the tetraspanin family of cell surface proteins, which play a role in cell adhesion, motility, and signal transduction.

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CD9-APC clone HI9A (3 citations) APC)–conjugated anti-CD9 (clone H19a, (2 citations)

5 protocols using cd9 apc

Multiparametric Immune Cell Analysis

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Cells were stained using the following fluorophore conjugated anti-mouse antibodies: WGA-488 (Invitrogen), Ly6G-FitC (clone 1A8, BioLegend), Ly6G-BV510 (clone 1A8, BioLegend), Ly6G-PE (clone 1A8, BioLegend), CD11c-BV605 (clone N418, BioLegend), F4/80-FitC (clone BM8, BioLegend), F4/80-PE (clone BM8, BioLegend), CD11b-PE (clone M1/70, BioLegend), Ly6C-PE Cy7 (clone HK1.4, BioLegend), CD63-APC (clone NVG-2, BioLegend), CD9-APC (clone MZ3, BioLegend), IL-1α-PE (clone ALF-161, BioLegend), and IL-1β-APC (clone NJTEN3, ThermoFisher Scientific). Antibodies were diluted in wash buffer (PBS with 1% BSA and 2 mM EDTA). Cells were stained for 20 minutes at 4°C, washed with wash buffer, fixed for 15 minutes in BD Biosciences cytofix/cytoperm, and permeabilization prior to intracellular staining. ACEA Novocyte was used for flow cytometry, and Novoexpress software was used for subsequent analysis. AMNIS ImageStream was used for imaging flow cytometry and the AMNIS IDEAS software was used to calculate the colocalization coefficient.

Ratitong B., Marshall M, & Pearlman E. (2021). β-Glucan-stimulated neutrophil secretion of IL-1α is independent of GSDMD and mediated through extracellular vesicles. Cell reports, 35(7), 109139.

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Immune Phenotyping of Extracellular Vesicles

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The immune phenotype of EVs was analyzed using flow cytometry (BD FACS Aria III, BD Bioscience, East Rutherford, NJ, USA) by immunostaining with the CD49e-PE (Sony, Tokyo, Japan), CD63-FITC (Biolegend, San Diego, CA, USA), Sca1-APC/Cy7 (BioLegend), CD45-PE/Cy7 (BioLegend), CD9-APC (Biolegend) and CD44-APC/Cy7 (BioLegend) antibodies.

Kostennikov A., Kabdesh I., Sabirov D., Timofeeva A., Rogozhin A., Shulman I., Rizvanov A, & Mukhamedshina Y. (2022). A Comparative Study of Mesenchymal Stem Cell-Derived Extracellular Vesicles’ Local and Systemic Dose-Dependent Administration in Rat Spinal Cord Injury. Biology, 11(12), 1853.

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Microglia Immunophenotyping by Flow Cytometry

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Microglia were treated with FC block (1:100; BD Biosciences 553142) 5 min in FACS buffer (DPBS, 2% BSA, 50 μM EGTA) before adding live/dead stain (1:100; Biolegend 423113), CD9-APC (1:100; Biolegend 312108) or CXCR4-APC (1:50; Biolegend 306510) for 30 min at 4 °C in the dark. Samples were washed 3× in FACS buffer. 100,000 events per sample were collected on BD LSRFortessa with FACSDiva 9.0 and analyzed in FlowJo 10.7.1. Gates were drawn on fluorescence minus one (FMO) controls.

McQuade A., Kang Y.J., Hasselmann J., Jairaman A., Sotelo A., Coburn M., Shabestari S.K., Chadarevian J.P., Fote G., Tu C.H., Danhash E., Silva J., Martinez E., Cotman C., Prieto G.A., Thompson L.M., Steffan J.S., Smith I., Davtyan H., Cahalan M., Cho H, & Blurton-Jones M. (2020). Gene expression and functional deficits underlie TREM2-knockout microglia responses in human models of Alzheimer’s disease. Nature Communications, 11, 5370.

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Multiparametric Immune Cell Analysis

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Ratitong B., Marshall M, & Pearlman E. (2021). β-Glucan-stimulated neutrophil secretion of IL-1α is independent of GSDMD and mediated through extracellular vesicles. Cell reports, 35(7), 109139.

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Comprehensive Characterization of Mesenchymal Cell Extracellular Vesicles

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The immune phenotype of CIMVs-MSCs and MSCs-derived EVs was characterized by immunostaining with the following monoclonal antibodies: Sca1-APC/Cy7 (BioLegend, USA), CD49e-PE (1119525; Sony, USA), CD44-APC/Cy7 (BioLegend, USA), CD45-PE/Cy7 (BioLegend, USA), CD9-APC (Biolegend, USA), CD63-FITC (Biolegend, USA). CIMVs and EVs were analyzed by flow cytometry (BD FACS Aria III. BD Bioscience, USA), the 405 nm laser was used for better distinguish CIMVs and and EVs from debris.

Gomzikova M.O., Aimaletdinov A.M., Bondar O.V., Starostina I.G., Gorshkova N.V., Neustroeva O.A., Kletukhina S.K., Kurbangaleeva S.V., Vorobev V.V., Garanina E.E., Persson J.L., Jeyapalan J., Mongan N.P., Khaiboullina S.F, & Rizvanov A.A. (2020). Immunosuppressive properties of cytochalasin B-induced membrane vesicles of mesenchymal stem cells: comparing with extracellular vesicles derived from mesenchymal stem cells. Scientific Reports, 10, 10740.

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