Human CD8 T cells were enriched from peripheral blood mononuclear cells (PBMCs) and, after sorting, were stained with CD4-FITC (Clone: OKT4) and CD8-APC (Clone: SK1). Analysis was performed by gating on live cells (Live+). Flow cytometric analysis revealed that the purity of the sorted Human CD8 T cells was 90.4%.
Human CD8 T Cell Isolation Kit (Column-Free)
Human CD8 T Cell Isolation Kit (Column-Free)
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Product Details
Product Details
Product Specification
| Format |
1 mL Human CD8 T Cell Biotin-Antibody Cocktail 1mL Streptavidin Beads |
| Capacity | Suitable for a total cell count of 1×10⁹, and can perform up to 100 sorting runs. |
| Principle of Separation | In this negative selection process, non-CD8 T cells are labeled with biotin-conjugated antibodies, and the cells are then incubated with streptavidin-nanomagnetic beads. As a result, the non-target cells are marked by the antibody‑bead complex, while the target CD8 T cells remain unlabeled. Subsequently, the cell suspension is transferred to a polystyrene tube and placed within the magnetic field of the sorter. Under the influence of the magnetic field, the magnetically labeled non-target cells adhere to the tube wall, whereas the unlabeled CD8 T cells remain in the supernatant. This supernatant constitutes the T-cell‑enriched fraction after removal of the non-target cells. |
| Reactivity | Human |
| Magnetic Bead Size | 150 nm |
| Separation Method | Column-Free |
| Selection Strategy | Negative Selection |
| Stability & Storage | Store protected from light at 2–8 °C; do not freeze. |
Background
Human CD8 T lymphocytes are the core effector cells of adaptive cellular immunity in the human body, playing a crucial role in host anti-infectious immunity, tumor immune surveillance, and immune homeostasis regulation. Due to their precise antigen specificity, potent cytotoxic function, and immunological memory characteristics, human CD8 T cells are central research subjects in fields such as immunological mechanism studies, anti-infectious vaccine development, tumor immunotherapy (e.g., CAR-T cell therapy), and immune function assessment, possessing significant scientific research and clinical application value. This kit employs a column-based negative selection approach: non-CD8 T cells are labeled with biotin-conjugated antibodies, and magnetic bead separation is used to remove non-target cells, thereby enriching highly pure, high‑activity CD8 T cells. This method avoids the potential impact of direct labeling of CD8 T cells on cellular function and is suitable for subsequent experimental studies, including cell culture and functional assays.
Protocol
Note: This kit does not include sorting buffer; please provide it separately. The corresponding Stater product number isS0D3018
Steps |
Operating Instructions |
Dosage and timing |
1 |
Peripheral blood mononuclear cells (PBMCs) are isolated from peripheral blood by density-gradient centrifugation using lymphocyte separation medium, or cryopreserved PBMCs are directly thawed and resuspended, followed by cell counting. Note: It is recommended to count cells after staining with AO/PI; PBMCs can be centrifuged at 400g for 7 minutes. |
Prepare the sample |
2 |
Resuspend 1×10 in 100 μL of MagSep Separation Buffer.sevencell; Note:Reagents are adjusted proportionally based on the sample volume; a pre-sorting sample is retained for assessing the proportion of human CD8 T cells. |
1×10sevencells/100 μL |
3 |
Add 10 μL of the Human CD8 T cell Biotin Antibody Cocktail to the sample and gently mix. Note:In this step, gently pipette up and down 2–3 times to mix thoroughly; |
10 μL/100 μL |
4 |
Incubate the antibodies in the cocktail with the cells at room temperature for 10 minutes; |
Incubate at room temperature for 10 minutes |
5 |
Shake for 5–30 seconds to mix the Streptavidin beads; |
Shake for 5–30 seconds |
6 |
Add 10 μL of Streptavidin beads to the sample; |
10 μL/100 μL |
7 |
Gently mix the magnetic beads and cells, and incubate at room temperature for 5 minutes; Note:For mixing in this step, you can use a pipette to gently blow the mixture 2–3 times; During this step, the magnetic beads may settle at the bottom; you can mix them again after about 2.5 minutes; |
Incubate at room temperature for 5 minutes |
8 |
Add MagSep Separation Buffer to the sample and immediately place it on the magnetic stand to prevent the magnetic beads from settling excessively; |
Add the total volume to 2.5 mL (5 mL flow cytometry tube) Add the total volume to 7.5 mL (15 mL centrifuge tube) |
9 |
Place the sample on the magnetic stand so that the magnetic beads adhere to the tube wall; |
Adsorb using a single-well magnetic stand (3 min) Adsorb using a multi-well magnetic stand (5 min) |
10 |
Tilt the magnetic stand and pour the sample into a new collection tube to collect the sample; Note:The sorted cells can be centrifuged at 400g for 7 minutes, suitable for subsequent culture and analysis. |
Cell sorting successful |
|
Note:For 0.1–1 × 10^8 cells (0.1–1 mL), it is recommended to use a 5 mL single-well magnetic separator (Starter EasyIso Separator) with a total volume of 2.5 mL; 2–5 × 10^8 cells (2–5 mL) are recommended to be adsorbed using a 15 mL row-type magnetic separator (Starter EasyEights EasyIso Separator), with a total volume of 7.5 mL; | ||
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Validation Data
