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Human T Cell Isolation Kit (Column-Based)

Human T Cell Isolation Kit (Column-Based)

Catalog Number: S0K1004 Reactivity: Human Conjugation: Brand: Starter
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Product Details

Product Specification


Format

1 mL Human T Cell Biotin-Antibody Cocktail

1 mL 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-T cells are labeled with biotin-conjugated antibodies, and the cells are then incubated with streptavidin-coated nanomagnetic beads. As a result, the non-target cells are marked by the antibody‑bead complex, while the target T cells remain unlabeled. Subsequently, the cell suspension is transferred to a separation column; under the influence of a magnetic field, the magnetically labeled non-target cells are retained in the column, whereas the unlabeled T cells elute into a collection tube under gravity. The resulting supernatant represents the T-cell‑enriched fraction after removal of the non-target cells.
Reactivity Human
Magnetic Bead Size 20 nm
Separation Method Column-Based
Selection Strategy Negative Selection
Stability & Storage

Store protected from light at 2–8 °C; do not freeze.

Background

Human T cells are key immune cells in the immune system, playing a central role in immune responses, immune regulation, and defense against infection and tumors. This kit employs a column-based negative selection approach: non-T cells are labeled with biotin-conjugated antibodies, and unwanted cells are removed using magnetic bead separation, thereby enriching highly pure, high‑activity T cells. This method avoids the potential impact of direct labeling on T cell function and is suitable for downstream experimental studies such as cell culture and functional assays.

Protocol

Note: This kit does not include the separation buffer; you need to prepare it yourself. The corresponding Stater catalog number isS0D3018

TakingL Separation Columnas an example

Steps

Operating instructions

Dosage and time

Cell processing and labeling

1

Isolate PBMCs from peripheral blood using density gradient centrifugation with lymphocyte separation solution, or directly thaw cryopreserved PBMC cells, and then perform cell counting.

Note: It is recommended to count cells after staining with AO/PI; PBMCs can be centrifuged at 400 g for 7 minutes.

Prepare the sample

2

Resuspend 1×10^7 cells in 100 μL of MagSep Separation Buffer.7cells;

Note:Adjust reagents proportionally according to the sample volume; set aside a pre-sorting sample to determine the proportion of human T cells.

1×10^77cells/100 μL

3

Add 10 μL of Human 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

Mix wellStreptavidin Beads, add 10 μLStreptavidin Beadsinto the sample;

10 μL/100 μL

6

Gently mix the magnetic beads and cells, and incubate at room temperature for 5 minutes;

Incubate at room temperature for 5 minutes

Cell sorting

7

Place it in the column-type magnetic separator,L Separation Column, add 3 mL of MagSep Separation Buffer to rinse the separation column;

Rinse the separation column

8

Add the cells to the separation column and collect the target cells using a collection tube;

Note:The cell volume should be at least 500 μL; if insufficient, add MagSep Separation Buffer to make up the volume. The cells that drip down under gravity are the target cells.

Collect the target cells

9

Add 3 mL of MagSep Separation Buffer and collect the target cells.

Note:In this step, elute the unlabeled target cells from the separation column to increase the yield.

Collect the target cells

Notes: Pay attention to the maximum capacity of the separation column.The maximum number of labeled cells in an S Separation Column is1×107;The maximum number of labeled cells in an L Separation Column is1×108;


Picture

Validation Data

Human T cells were enriched from peripheral blood mononuclear cells (PBMCs). After fluorescent staining with CD45-FITC (SDT Cat: S0B8067) and CD3-APC (SDT Cat: S0B5722), live cells (Live+) were gated for analysis. Flow cytometry results showed that the purity of the sorted Human T cells was 97.8%.