The International Society for Cell Therapy previously suggested the following minimal criteria to define human MSCs [18] (link): expression of CD105, CD73, and CD90, and no expression of CD45, CD34, CD14, CD11b, CD79α, CD19, or HLA-DR surface molecules. Cells were harvested by treatment with 0.05% trypsin-EDTA (Sigma-Aldrich, St Louis, MO) for 3 min at 37°C, recovered by centrifugation at 400× g for 5 min, washed twice in ice-cold PBS containing 2% FBS, and re-suspended at 1×105 cells/antibody test. The expression of specific MSC markers was assessed using the following antibodies: CD14-FITC, CD31-PE, CD105-FITC, CD34-PE, CD45-PE, CD73-PE, and CD90-APC (all purchased from Abcam, Cambridge, UK). Negative control staining was performed using FITC/PE/APC-conjugated mouse IgG1 isotype antibodies (Abcam). After incubation for 30 min at room temperature in the dark, the cells were washed with PBS, resuspended in 100 µL PBS, and analyzed by a MACSQuant flow cytometer (Miltenyi Biotec, Gladbach, Germany).
Cd45 pe
Manufactured by Abcam
Sourced in United Kingdom
CD45-PE is a conjugated antibody used for the identification and enumeration of cells expressing the CD45 antigen. CD45 is a transmembrane protein tyrosine phosphatase that is expressed on the surface of all nucleated hematopoietic cells. The PE (Phycoerythrin) fluorescent dye is conjugated to the CD45 antibody, allowing for detection and analysis of CD45-positive cells by flow cytometry.
Automatically generated - may contain errors
Lab products found in correlation
Trypsin edta, by Merck Group (1 mentions)
Macsquant flow cytometer, by Miltenyi Biotec (1 mentions)
Cd105 fitc, by Abcam (1 mentions)
Cd34 pe, by Abcam (1 mentions)
Cd73 pe, by Abcam (1 mentions)
Cd90 apc, by Abcam (1 mentions)
Cd31 pe, by Abcam (1 mentions)
Cd105 pe, by Abcam (1 mentions)
Cd90 pe, by Abcam (1 mentions)
Cd11b pe, by Abcam (1 mentions)
Cd29 pe, by Abcam (1 mentions)
Cd34 fitc, by Abcam (1 mentions)
2 protocols using cd45 pe
1
Minimal Criteria for Human MSCs
2
Flow Cytometry Analysis of Cultured Cells
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The third-generation cultured cells were digested with 0.25% trypsin for 5 minutes. After centrifugation at 1,000 rpm for 10 minutes, the cells were washed twice with PBS, and then the cells were resuspended with PBS to prepare cell suspensions. The cell concentration was adjusted to 2×106/mL. The cells were divided into flow centrifuge tubes at 500 µL per tube, and CD11B-PE (Abcam Cambridge, UK), CD29-PE (Abcam, UK), CD-34-FITC (Abcam, UK), CD45-PE (Abcam, UK), CD90-PE (Abcam, UK), CD105 PE (Abcam, UK), and the isotype control (Abcam, UK) were added. The amount of each added antibody was 10 µL. After incubation at 4 °C for 30 minutes, cells were washed twice with PBS, then 4% paraformaldehyde (Guangzhou Jibeisi Biological Technology Co., Ltd.) at 200 µL was added to each tube to resuspend the cells. A filter membrane was used to filter cell masses to prevent blockage of the flow cytometer. Cells were then fed into the flow cytometer for identification.
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