UC-MSCs were harvested and placed into tubes at a density of 1×105 cells/tube. Cells were washed twice in phosphate-buffered saline (PBS) by centrifugation at 180 × g for 5 min at room temperature, and labeled with the following antibodies (10 µl/tube) purchased from BD Pharmingen (San Diego, CA, USA) for 1 h at 4°C: Mouse anti-cluster of differentiation (CD) 11b-phycoerythrin (PE; catalog no. 555388), mouse anti-CD19-PE (catalog no. 555413), mouse anti-CD31-fluorescein isothiocyanate (FITC; catalog no. 555445), mouse anti-CD34-allophycocyanin (APC; catalog no. 555824), mouse anti-CD44-PE (catalog no. 555479), mouse anti-CD45-FITC (catalog no. 555482), mouse anti-CD73-PE (catalog no. 550257), mouse anti-CD90-FITC (catalog no. 555595), mouse anti-CD105-PE (catalog no. 560839), mouse anti-CD106-PE (catalog no. 555647), mouse anti-CD166-PE (catalog no. 559263) and human leukocyte antigen D related (HLA-DR)-FITC (catalog no. 555811). FITC-(catalog no. 555748), APC-(catalog no. 555751) or PE-conjugated (catalog no. 555749) isotype control antibodies served as controls. Following incubation, UC-MSCs were washed with PBS, resuspended in 2% paraformaldehyde and analyzed on a BD™ LSR II flow cytometer (BD Biosciences) using BD FACSDiva™ software version 6.1.3 (BD Biosciences).
Mouse anti cd106 pe
Manufactured by BD
Sourced in United States
Mouse anti-CD106-PE is a flow cytometry reagent used for the detection and quantification of CD106, also known as VCAM-1, on the surface of cells. It consists of an anti-CD106 monoclonal antibody conjugated to the fluorescent dye phycoerythrin (PE).
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3 protocols using mouse anti cd106 pe
1
Phenotypic Characterization of UC-MSCs
2
Flow Cytometry Analysis of Second-Generation BMSCs
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The density of second-generation BMSCs was adjusted to 2 × 107 cells/mL. The control group received 50 µL of buffer, while the single-label group received 5 µL of hamster anti-CD29-AF647 (BD Biosciences, Franklin Lakes, NJ, USA), mouse anti-CD90-PECyTM7 (BD Biosciences), mouse anti-CD106-PE (BD Biosciences), mouse anti-CD11b-V450 (BD Biosciences), or mouse anti-CD45-FITC (BD Biosciences) to each branch-flow sampling tube, after which 45 µL of buffer was added to each tube. The multicolor group received 5 µL of each antibody into a one-branch-flow sampling tube, and then, 25 µL of buffer was added. Next, 50 µL of cell suspension was added to each flow tube, the tubes were incubated at room temperature for 30 min and washed twice with staining buffer, and then, 500 µL of buffer was added to each tube for detection by flow cytometry (Beckman Coulter Life Sciences, Brea, CA, USA).
3
Multiparametric Flow Cytometry of Mouse Leukocytes
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First, 5 μL of mouse anti-CD90-PECyTM7, mouse anti-CD106-PE, mouse anti-CD11b-V450, and mouse anti-CD45-fluorescein isothiocyanate (FITC) (BD Biosciences, Franklin Lakes, NJ, USA) was added into an flow cytometry (FCM) tube, followed by the addition of 50 μL cell suspension (2 × 107/mL). The mixture was incubated at room temperature (RT) in the dark for 30 min, and the contents of each tube were then washed twice using a standing buffer. Then, 500 μL staining buffer was added to resuspend the cells, and FCM (Beckman Coulter Life Sciences, Brea, CA, USA) was used to detect expression levels of CD11b, CD45, CD90, and CD106.
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