Small EVs (5 × 1010) were stained with CellMask Deep Red plasma membrane stain for 30 min at 37°C. The stained small EVs were placed in three groups of equal numbers and incubated with PBS only, a human PE-conjugated anti-CD63 antibody (R&D Systems; Cat. #IC5048P-025, 1:10), and a human PE-conjugated anti-VEGF-A antibody (R&D Systems; Cat. #IC2931P, 1:10), respectively. After 1 hr of incubation at room temperature, the samples were diluted with 3 mL of PBS and centrifuged for 3 hr at 100,000 × g and 4°C. The pellets were re-suspended in 50 μL of PBS and immediately analyzed using an Amnis Image StreamX MKII flow cytometer (Luminex). For single-color control samples, 20 μL of OneComp eBeads Compensation Beads (Thermo Fisher Scientific) was incubated with the PE-conjugated anti-CD63 antibody for 20 min at room temperature. The beads were then diluted with 1 mL of PBS and centrifuged for 5 min at 1500 rpm and 4°C. The PE-conjugated bead pellet was eventually re-suspended in 50 μL of PBS. The samples exposed to PBS only, PBS with CellMask Deep Red plasma membrane stain, or PBS with PE-conjugated anti-VEGF antibodies served as negative controls. Experiments were performed three times independently.
Human pe conjugated anti cd63 antibody
Manufactured by R&D Systems
The Human PE-conjugated anti-CD63 antibody is a laboratory reagent designed for the detection and analysis of CD63 protein expression in human samples. The antibody is conjugated with the fluorescent dye Phycoerythrin (PE), allowing for the visualization and quantification of CD63-positive cells using flow cytometry or other fluorescence-based techniques.
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2 protocols using human pe conjugated anti cd63 antibody
1
Characterization of Small Extracellular Vesicles
2
Characterization of Small Extracellular Vesicles
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Small EVs (5 × 1010) were stained with CellMask Deep Red plasma membrane stain for 30 min at 37°C. The stained small EVs were placed in three groups of equal numbers and incubated with PBS only, a human PE-conjugated anti-CD63 antibody (R&D Systems; Cat. #IC5048P-025, 1:10), and a human PE-conjugated anti-VEGF-A antibody (R&D Systems; Cat. #IC2931P, 1:10), respectively. After 1 hr of incubation at room temperature, the samples were diluted with 3 mL of PBS and centrifuged for 3 hr at 100,000 × g and 4°C. The pellets were re-suspended in 50 μL of PBS and immediately analyzed using an Amnis Image StreamX MKII flow cytometer (Luminex). For single-color control samples, 20 μL of OneComp eBeads Compensation Beads (Thermo Fisher Scientific) was incubated with the PE-conjugated anti-CD63 antibody for 20 min at room temperature. The beads were then diluted with 1 mL of PBS and centrifuged for 5 min at 1500 rpm and 4°C. The PE-conjugated bead pellet was eventually re-suspended in 50 μL of PBS. The samples exposed to PBS only, PBS with CellMask Deep Red plasma membrane stain, or PBS with PE-conjugated anti-VEGF antibodies served as negative controls. Experiments were performed three times independently.
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