BeaverBeads® Human CD3⁺ Cell Isolation Kit (PositiveSelection)
The Human CD3⁺ Cell Isolation Kit (PositiveSelection) is designed for isolating CD3⁺ cells from human peripheral blood mononuclear cells (PBMCs). The principle involves labeling CD3+ cells with a CD3 Capture Antibody, followed by capturing the target cells using Releasable BeaverBeads®. The magnetic beads are then dissociated from the cell surface using BeaverBeads® Release Buffer, resulting in CD3+ cells free of magnetic bead labeling. The sorted CD3+ cells can be used in downstream molecular biology and cell biology experiments.
Product Features
Streamlined Workflow: No separation column required; isolation is completed using a magnetic separator.
High Purity: Achieves highgrade separation purity, with postisolation CD3⁺ cell purity up to 95%.
High Viability: Cell viability remains unaffected throughout the procedure, yielding beadfree CD3⁺ cells with intact biological functions.
Performance Data
1. Cell Purity Analysis
CD3⁺ cells were isolated from human PBMCs with the BeaverBeads® Human CD3⁺ Cell Isolation Kit (PositiveSelection). Before and after isolation, Cells were stained with PEconjugated antihuman CD3 antibody (Clone: OKT3) and analysed by flow cytometry. As shown in Figure 1, CD3⁺ cells accounted for 55.0 % in the starting PBMC sample, and reached a purity of 94.1 % (93.9±0.9 %) after isolation.
Figure 1. Flowcytometric analysis of CD3⁺ cells
2. Cell Viability Analysis
CD3⁺ cells were isolated from human PBMCs using the BeaverBeads® Human CD3⁺ Cell Isolation Kit (Positive selection). Cell viability was assessed by flow cytometry using 7-AAD (PerCP) staining before and after isolation. As shown in Figure 2, the viability of input cells was 98.9 %, and postisolation cell viability was 99.5 %.
Figure 2. Flowcytometric analysis of CD3⁺ cells viability
3. Cellular Functional Assay
CD3⁺ cells were isolated from human PBMCs with the BeaverBeads® Human CD3⁺ Cell Isolation Kit (PositiveSelection). Purified CD3⁺ cells were cocultured with BeaverBeads® Human CD3/CD28 TCell Activation Beads (Cat.No. 70910). After 24 hours, the proportion of CD25⁺CD69⁺ Tcells was determined by flow cytometry.
As illustrated in Figure 3, CD3⁺ cells isolated by this kit can be efficiently activated by Tcellactivation beads. The percentage of CD25⁺CD69⁺ Tcells shifted from 7.1 % in the unstimulated group to 52.7 % upon stimulation.
Figure 3. Flow cytometric analysis of CD25 and CD69 expression on CD3⁺ cells.
