LIVE/DEAD® Cell Imaging Kit (R37601 Thermo Scientific®) was used following the manufacturer’s instructions. This viability staining was performed on the complete and living 3D structure. For hematopoietic antigens, 3D-MS serial slides were incubated at room temperature with primary antibodies (Anti-CD34 APC clone 581 Thermo Scientific®, anti-Ki67 clone 4A1 Thermo Scientific®, anti-CD133 clone AC133 Miltenyi Biotec®, anti-CD33 clone WM53ThermoScientific® and anti-CD7 clone MEM-186, Thermo Scientific®). Alexa Fluor 514 goat anti-mouse IgG (Thermo Scientific®) was used as a secondary antibody for CD133, CD33 or CD7 detection. Anti-α-tubulin (Alexa Fluor 488 clone B512, Invitrogen®) was used for cytoskeletal labeling, and DAPI (4ʹ,6-diamidino-2-phenylindole, ref D1306, Invitrogen®) was used for nuclei identification. Slide-mounted 3D structure sections were sealed with 10 μL of ProLong Gold antifade reagent (Invitrogen®) and covered with coverslips.
Anti cd34 apc clone 581
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Anti-CD34 APC clone 581 is a fluorescently-labeled antibody that binds to the CD34 antigen expressed on the surface of certain cell types. This product is intended for use in flow cytometry applications to identify and quantify CD34-positive cells.
Automatically generated - may contain errors
Lab products found in correlation
Prolong gold antifade reagent, by Thermo Fisher Scientific (1 mentions)
Live dead cell imaging kit, by Thermo Fisher Scientific (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions)
Anti cd33 clone wm53, by Thermo Fisher Scientific (1 mentions)
Facsaria iiu cytometer, by BD (1 mentions)
Anti cd45 percp clone 2d1, by BD (1 mentions)
2 protocols using anti cd34 apc clone 581
1
3D Immunofluorescence Staining of Hematopoietic Cells
2
Isolation and Characterization of MSCs
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To obtain MSC, reaming samples were collected from the intramedullary canal of the femoral head from 4 patients undergoing prosthetic hip replacement who attended the Department of Orthopedics and Traumatology of Hospital Universitario San Ignacio (Bogotá, Colombia). The sample donors participated voluntarily and accepted the informed consent previously reviewed and approved by the independent ethics committees (IECs) of the Hospital Universitario San Ignacio (Approval: 04/2020) in the ordinary session of 12 March 2020. MSCs were isolated and cultured according to protocols previously published by our group [44 (link),47 (link),48 (link)] and ISO 24651 international standard. MSC phenotype was confirmed by evaluating antigens as: CD34 (anti CD34-APC, clone 581, Thermo Scientific®, Waltham, MA, USA), CD45 (anti CD45-PerCP, clone 2D1, BD Biosciences®), CD73 (anti CD73-FITC, clone AD2, BD Pharmingen™, San Diego, CA, USA) and CD105 (anti CD105-PE, clone SN6, Thermo Scientific®) by flow cytometry using the FACSAria II-U cytometer (BD Biosciences®) and the data were analyzed using FlowJo 10.8.1 Software (Tree star, Ashland, OR, USA).
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