Cd10 pe

Manufactured by BioLegend

Sourced in United States, United Kingdom

CD10-PE is a fluorochrome-conjugated monoclonal antibody that binds to the CD10 cell surface antigen. CD10 is a cell surface metalloprotease enzyme that is expressed on a variety of cell types, including B-cell precursors, germinal center B cells, and granulocytes. The PE (phycoerythrin) fluorochrome allows for the detection and analysis of CD10-positive cells using flow cytometry.

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Close spelling variants of this product

CD10 PE-Cy7 (7 citations) CD10-PE-Dazzle594 (4 citations) Anti-human CD10-PE-Cy7 (3 citations) Anti-CD10-PE (2 citations) PE-Cy7-anti-CD10 (2 citations) PE-Cy7 mouse anti-human CD10 (2 citations)

5 protocols using cd10 pe

Isolation and Characterization of Murine MSCs

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MSCs were isolated from subcutaneous adipose tissue of the adult mice (8–9 weeks old). Adipose tissue were dissected into small pieces and digested with 0.2% collagenase II (Dia-M, Russia) in a shaker-incubator at 37 °C, 120 rpm for one hour. Suspended cells were pelleted (500 g for 5 min), washed once in PBS (PanEco, Russia) and re-suspended in DMEM (PanEco, Russia) supplemented with 10% fetal bovine serum (Gibco, UK) and 2 mM l-glutamine (PanEco, Russia). MSCs were maintained at 37 °C, 5% CO₂ with culture medium being replaced every 3 days. MSCs from passages 3 and 4 were used for the experiments.
MSCs were differentiated into three lineages: adipogenic, chondrogenic and osteogenic and it was confirmed by detection of lipid droplets (Oil Red dye staining), glycosaminoglycans and mucins (1% alcian blue staining) and calcium deposits (5% AgNO₃ staining), respectively. Immune phenotype was determined using monoclonal antibodies to CD90.2-PerCP (1301575; Sony, USA), CD44-APC/Cy7 (103028; BioLegend, USA), CD73-Alexa Fluor647 (127208; BioLegend, USA), CD49e-PE (1119525; Sony, USA), Sca1-APC/Cy7(108126; BioLegend, USA), CD29-PE (102208; BioLegend, USA), CD11b-PE/Cy7 (101216; BioLegend, USA), CD10-PE (312203; BioLegend, USA), CD45-PE/Cy7 (103114; BioLegend, USA). Expression of CD markers was analyzed by flow cytometry (BD FACS Aria III (BD Bioscience, USA)).

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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Multicolor Flow Cytometry for Cell Phenotyping

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CD227-FITC (BD Bioscience #559774, clone HMPV, 1:50), CD10-PE (BioLegend #312204, clone HI10a, 1:100), CD10-APC (BioLegend #312210, clone HI10a, 1:100), CD227-PE (BioLegend #355604, clone 16A, 1:100) were added to cells in media for 25 min on ice, washed in PBS and analyzed with a FACSCalibur (Becton Dickinson).

Lee J.K., Garbe J.C., Vrba L., Miyano M., Futscher B.W., Stampfer M.R, & LaBarge M.A. (2015). Age and the means of bypassing stasis influence the intrinsic subtype of immortalized human mammary epithelial cells. Frontiers in Cell and Developmental Biology, 3, 13.

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Immunophenotyping of Adherent Cells

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Subconfluent cells were trypsinized and fixed in 2% paraformaldehyde. Cells were blocked in FACS buffer (2.5% BSA, 1mM EDTA in 1XPBS) and stained with CD227-FITC (BD Biosciences, clone HMPV, 1:50) and CD10-PE (BioLegend, clone HI10a, 1:100) in FACS buffer for 25 min on ice protected from light, washed in PBS, and analyzed using FACS Calibur (Becton Dickinson). FlowJo X was used to for computer analysis.

Lee J.K., Bloom J., Zubeldia-Plazaola A., Garbe J.C., Stampfer M.R, & LaBarge M.A. (2018). Different culture media modulate growth, heterogeneity, and senescence in human mammary epithelial cell cultures. PLoS ONE, 13(10), e0204645.

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Scaffold Cell Composition and Viability

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To assess the cellular composition and viability in the scaffolds, cells were stained with the DNA-intercalating viability dye 7-AAD, and the antibodies CD34-ApCCy7, CD38-APC, CD73-FITC and CD10-PE (all from BioLegend, London, UK). Briefly, cells were washed once (300×g, 5 min, 4 °C), resuspended in PBS-0.5% BSA and placed on ice. Antibody cocktails (4 colour and fluorescence minus one (FMO)) were prepared in the dark, 250 μL cell samples were transferred to a 96 well round bottom ULA plate (Corning) and stained with antibody cocktails at 4 °C for 30 min in the dark. Plates were washed twice (300×g, 5 min, 4 °C) and pellets resuspended in 250 μL fresh PBS-0.5% BSA before acquisition on the Guava easyCyte 8HT flow cytometer (Millipore, Burlington, MA, USA). Guava single stained beads were used as antibody compensation controls following manufacturer’s instructions (AbC total Antibody Compensation Bead Kit; Thermo Fischer Scientific, Waltham, MA, USA). Freeze-killed 3D model-harvested cells were used for the 7-AAD compensation control (stain dead cells), vehicle-treated cells were used for the unstained control. Data were analysed using FlowJo 10 (FlowJo LLC, Ashland, OR, USA).

David R., Gee S., Khan K., Wilson A, & Doherty A. (2021). Three dimensional and microphysiological bone marrow models detect in vivo positive compounds. Scientific Reports, 11, 21959.

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Flow Cytometry Analysis of B Cell Subsets

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Cells were stained for flow cytometry analysis using the following reactive antibodies and reagents: CD38-AF700 (HIT2), CD27-APC (O323), CD21-PcP-Cy5.5 (Bu32), CD24-APC-Cy7 (ML5), CD10-PE (HI10a), IgM-Pacific Blue (MHM-88), CD5-BV785 (UCHT2), CD3-BV510 (OKT3), CD14-BV510 (M5E2), CD16-BV510 (3G8), CD19-BUV395 (SJ25C1), IgG-PE-Cy7 (M131G05), and IgD-FITC (Ia6-2) (BioLegend, BD Biosciences, eBioscience, or Tonbo Biosciences). Jo-1 binding cells were identified through staining with GST-Jo-1 followed by detection with anti-GST-Texas Red (Abcam). Samples were acquired using a LSR II flow cytometer (BD Biosciences) and data were analyzed using FlowJo software (Tree Star). Flow cytometry samples identifying less than 20 JBC events were excluded from downstream analysis of B cell subset composition. Non-JBC subset analysis was still performed for these donors.

Young-Glazer J., Cisneros A I.I.I., Wilfong E.M., Smith S.A., Crofford L.J, & Bonami R.H. (2021). Jo-1 autoantigen-specific B cells are skewed towards distinct functional B cell subsets in anti-synthetase syndrome patients. Arthritis Research & Therapy, 23, 33.

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