Human CD3+ T Cell Sorting Kit CS-0004 for column-based T cell isolation | Cell Sorts™
CS-0004

Human CD3+ T Cell Sorting Kit

Column-Based Negative Human

Cell Sorts™ Human CD3+ T Cell Sorting Kit (CS-0004) enriches untouched CD3+ T cells from human PBMCs by column-based negative selection. A biotin-antibody cocktail labels non-target cells, and anti-biotin nanobeads retain them in a magnetized separation column. Unlabeled CD3+ T cells are recovered in the combined flow-through and wash without direct bead labeling.

Supplied technical documentation specifies 5-minute antibody and 10-minute nanobead incubations at 2–8°C. For 1×10⁷ cells, use 10 µL cocktail and 10 µL nanobeads; one kit supports up to 1×10⁹ total cells and 100 separations. Representative flow cytometry shows post-sort CD3+ purity near 98%, with recovery and viability evaluated in parallel. For research use only.

Sorting Purity
Representative result near 98%
Sorting Time
5 + 10 min incubations

Product Features of the Human CD3+ T Cell Sorting Kit

The Human CD3+ T Cell Sorting Kit delivers industry-leading performance for rapid CD3+ T Cell isolation from PBMCs via column-based negative selection.

Column-based negative selection — A biotin-antibody cocktail and anti-biotin nanobeads retain non-target cells in a magnetized column
Untouched target fraction — CD3+ T cells are collected in the combined flow-through and wash without direct bead labeling
Representative purity near 98% — The supplied technical documentation shows a post-sort CD3+ fraction close to 98% in representative flow-cytometry data
Defined reagent ratio — Use 10 µL antibody cocktail and 10 µL nanobeads per 1×10⁷ cells, with proportional scaling for larger inputs
Scalable column formats — Small and medium column workflows support up to 1×10⁸ and 1×10⁹ total cells, respectively
Controlled cold labeling — Perform labeling at 2–8°C with pre-cooled buffer, then use room-temperature buffer for column separation

The Human CD3+ T Cell Sorting Kit uses column-based negative selection to isolate untouched CD3+ T Cells from human PBMCs. To achieve these results, follow this simple 4-step protocol that takes just 5 + 10 min incubations from start to finish. No centrifugation and no lengthy incubations. Furthermore, this streamlined protocol ensures consistent results across experiments.

How the Human CD3+ T Cell Sorting Kit Works

Simple 4-step protocol for rapid cell isolation

1
CS-0004 step 1: prepare PBMCs, centrifuge at 300×g for 10 minutes, and resuspend 1×10⁷ cells in 80 µL buffer

Step 1: Sample Preparation

Prepare PBMC single-cell suspension. Resuspend 1×10⁷ cells in 80 μL cold buffer. Add FcR blocker.

2
CS-0004 step 2: add 10 µL biotin-antibody cocktail per 1×10⁷ cells and incubate for 5 minutes at 2–8°C

Step 2: Antibody Cocktail Labeling

Add 10 μL biotin antibody cocktail per 1×10⁷ cells. Incubate 4°C 5 min. Labels non-CD3⁺ cells; T cells untouched.

3
CS-0004 step 3: add 10 µL anti-biotin nanobeads per 1×10⁷ cells, incubate for 10 minutes, and adjust to 500 µL

Step 3: Magnetic Bead Binding

Add 10 μL anti-biotin magnetic nanobeads per 1×10⁷ cells. Incubate 4°C 10 min. Beads bind labeled non-target cells.

4
CS-0004 step 4: load a rinsed magnetic column and combine the flow-through with one column wash to recover CD3+ T cells

Step 4: Column Separation & Collection

Load onto pre-washed magnetic column. Non-target cells retained. Untouched CD3⁺ T cells flow through — ready for use.

Applications for the Human CD3+ T Cell Sorting Kit

This Human CD3+ T Cell Sorting Kit supports a wide range of downstream applications for CD3+ T Cell isolation and column-based negative selection workflows from human PBMCs.

Flow Cytometric Immunophenotyping — Prepare an enriched CD3+ T-cell fraction for laboratory-validated phenotype, purity, viability, and subset analysis T Cell Activation and Proliferation — Evaluate activation or proliferation after confirming that the recovered fraction meets assay-specific requirements Cytokine and Signaling Studies — Support controlled studies of T-cell signaling and cytokine responses using an untouched target fraction Cell Culture and Expansion — Establish research culture or expansion workflows after post-sort cell counting and viability assessment TCR and Single-Cell Analysis — Prepare input for TCR, transcriptomic, or single-cell workflows when platform-specific QC criteria are met Infectious-Disease and Vaccine Research — Investigate human T-cell responses in appropriately controlled research models Tumor Immunology and Cell-Therapy Research — Support exploratory T-cell studies after laboratory-specific functional qualification Drug Discovery — Evaluate candidate immunomodulators in validated human T-cell assays

In addition, the Human CD3+ T Cell Sorting Kit is compatible with a wide range of downstream applications, making it a versatile tool for your immunology research workflow.

Kit Components

Each Human CD3+ T Cell Sorting Kit contains everything needed for high-purity column-based negative selection from human PBMCs.

  • Human CD3+ T Cell Biotin-Antibody Cocktail — 100 tests
  • Anti-Biotin Nanobeads — 100 tests

Storage & Shelf Life

Storage
Store at 2–8°C protected from light; do not freeze nanobeads
Shelf Life
Refer to the product label

Human CD3+ T Cell Sorting Kit — Frequently Asked Questions

Everything you need to know about the CS-0004 Human CD3+ T Cell Sorting Kit

Click each question to expand the answer

CS-0004 is intended for enriching untouched human CD3+ T cells from PBMCs prepared from peripheral blood by density-gradient separation. The supplied workflow includes washing the PBMCs and performing red-blood-cell lysis before labeling.
A biotin-conjugated antibody cocktail labels non-CD3+ cells, and anti-biotin nanobeads bind that labeled population. A compatible magnetized column retains the non-target cells while the unlabeled CD3+ T-cell fraction is recovered in the flow-through and column wash.
Representative flow-cytometry data in the supplied documentation show a post-sort CD3+ fraction close to 98%. Recovery and viability are also evaluated in the representative dataset. These are representative results rather than guaranteed specifications; donor biology, PBMC quality, cell counting, and protocol execution affect performance.
The supplied documentation does not state a validated total end-to-end processing time. It specifies a 5-minute antibody-cocktail incubation, a 10-minute nanobead incubation, a 10-minute sample centrifugation, and the required column rinsing, loading, flow-through, and wash steps.
For 1×10⁷ total cells, resuspend in 80 µL separation buffer and add 10 µL biotin-antibody cocktail plus a self-provided receptor blocker. After 5 minutes at 2–8°C, add 10 µL anti-biotin nanobeads and incubate for 10 minutes. Use the stated minimum volumes for smaller samples and scale all volumes proportionally for larger inputs.
The documented user-prepared separation buffer is PBS at pH 7.2 with 0.5% BSA and 2 mM EDTA. Keep cells and labeling buffer at 2–8°C, but do not place cells directly into ice because excessive cooling may promote nonspecific binding or apoptosis. Return the buffer to room temperature for column separation to reduce bubble formation.
The documented small column supports up to 1×10⁷ labeled cells and 1×10⁸ total cells. The medium column supports up to 1×10⁸ labeled cells and 1×10⁹ total cells. Use a compatible magnet, do not exceed column capacity, avoid visible bubbles, and do not reuse the column.
Rinse the small column with 500 µL buffer or the medium column with 3 mL buffer, load the labeled suspension, and collect the complete flow-through. Wash once with the same format-specific volume and combine the flow-through and wash; the combined fraction contains the enriched CD3+ T cells.
Use a validated flow-cytometry panel that includes CD3 and an appropriate viability marker. Report post-sort CD3+ purity, viable-cell recovery, total yield, and—when relevant—CD4/CD8 subset composition. The supplied documentation describes optional CD3 staining as a purity check.
Review PBMC viability and aggregation, red-blood-cell removal, cell counting, the 300×g centrifugation step, reagent scaling, 2–8°C incubation conditions, nanobead mixing, final cell concentration, column capacity, room-temperature buffer, column rinsing, bubble formation, and collection of both the initial flow-through and wash.

Related topics covered:

column-based negative selection CD3+ T Cell isolation CD3+ T Cell purity verification CD3+ T Cell sorting protocol column-based vs column-free single-cell analysis (scRNA-seq) CD3+ T Cell isolation markers PBMC CD3+ T Cell sorting

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