Human CD3+ T Cell Sorting Kit CS-0003 for column-free T cell isolation | Cell Sorts™
CS-0003

Human CD3+ T Cell Sorting Kit

Column-Free Negative Human

Cell Sorts™ Human CD3+ T Cell Sorting Kit (CS-0003) enriches untouched CD3+ T cells from human PBMCs by column-free negative selection. A biotin-antibody cocktail labels non-T cells, streptavidin nanobeads bind the labeled fraction, and a magnetic separator retains non-target cells on the tube wall. Unlabeled T cells are collected in a fresh tube without direct antibody or bead labeling.

Supplied technical documentation specifies 10-minute cocktail labeling, 5-minute nanobead binding, and 3–5 minutes of magnetic separation at room temperature. The documented workflow takes 18–20 minutes and supports 1×10⁸, 5×10⁸, or 1×10⁹ total cells. Representative flow cytometry showed 98.7% post-sort CD3+ purity and a proliferative response after stimulation. For research use only.

Sorting Purity
Representative result 98.7%
Sorting Time
18–20 min

Product Features of the Human CD3+ T Cell Sorting Kit

CS-0003 uses a documented column-free negative-selection workflow to enrich untouched human CD3+ T cells from density-gradient-prepared PBMCs.

Column-free negative selection — Depletes magnetically labeled non-T cells without a separation column
Untouched CD3+ target fraction — Antibodies and nanobeads do not directly label the recovered T cells
Representative 98.7% post-sort purity — A documented example result, not a guaranteed value for every sample
18–20 minute documented separation — 10-minute cocktail labeling, 5-minute nanobead binding, and 3–5-minute magnetic hold
Scalable kit formats — Options support up to 1×10⁸, 5×10⁸, or 1×10⁹ total starting cells
Tube-based collection — Recover the enriched fraction by controlled pour-off into a fresh tube

CS-0003 uses column-free negative selection to enrich untouched CD3+ T cells from human PBMCs. The documented 18–20 minute separation comprises 10-minute cocktail labeling, 5-minute nanobead binding, and a 3–5 minute magnetic hold; PBMC preparation, counting, optional washing, and QC are additional.

How the Human CD3+ T Cell Sorting Kit Works

Simple 4-step protocol for rapid cell isolation

1
Step 1 of CS-0003 protocol: prepare density-gradient PBMCs at no more than 1×10⁸ cells per mL in specified FACS buffer

Step 1: Prepare the PBMC Suspension

Prepare PBMCs by density-gradient separation, count viable cells, and resuspend at no more than 1×10⁸ cells/mL in FACS buffer (1× PBS, 2% FBS, 0.02% EDTA).

2
Step 2 of CS-0003 protocol: label non-T cells with isolation cocktail for 10 minutes at room temperature

Step 2: Label Non-Target Cells

Add 100 µL of isolation cocktail per mL of cell suspension (10 µL for 1×10⁷ cells in 100 µL), mix gently, and incubate for 10 minutes at room temperature. An optional wash may be used for selected patient PBMC samples.

3
Step 3 of CS-0003 protocol: bind labeled non-target cells with streptavidin nanobeads for 5 minutes at room temperature

Step 3: Bind Streptavidin Nanobeads

Mix the nanobeads for 5–30 seconds, add 100 µL per mL of cells, and incubate for 5 minutes at room temperature. Remix midway if bead settling is visible.

4
Step 4 of CS-0003 protocol: retain labeled cells magnetically and pour untouched CD3+ T cells into a fresh tube

Step 4: Magnetize and Collect CD3+ T Cells

Bring the sample to 2.5 mL in a 5 mL tube or 7.5 mL in a 15 mL tube. Magnetize for 3 minutes with a single-position separator or 5 minutes with a multi-position separator, then pour the untouched CD3+ fraction into a fresh tube.

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-free negative selection workflows from human PBMCs.

Flow Cytometric Immunophenotyping — Assess post-sort CD3+ purity, viability, and CD4/CD8 subset composition using a laboratory-validated panel T Cell Activation and Proliferation — Evaluate receptor-driven activation and proliferation in the enriched untouched T-cell fraction Cytokine and Signaling Studies — Investigate T-cell cytokine responses and intracellular signaling under controlled stimulation conditions Cell Culture and Expansion — Establish research culture or expansion workflows after confirming post-sort count and viability TCR and Single-Cell Analysis — Prepare input for TCR, transcriptomic, or single-cell workflows when platform-specific QC requirements are met Infectious-Disease and Vaccine Research — Study 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-free negative selection from human PBMCs.

  • Human CD3+ T Cell Isolation Cocktail
  • Streptavidin Nanobeads

Storage & Shelf Life

Storage
Store refrigerated at 4°C; do not freeze
Shelf Life
Refer to the product label

Human CD3+ T Cell Sorting Kit — Frequently Asked Questions

Everything you need to know about the CS-0003 Human CD3+ T Cell Sorting Kit

Click each question to expand the answer

CS-0003 is intended to enrich untouched CD3+ T cells from human PBMCs. Prepare PBMCs by density-gradient separation, determine viable-cell concentration, and produce a uniform single-cell suspension. The supplied documentation does not recommend routine red-blood-cell lysis for this workflow. For research use only.
A biotin-antibody cocktail labels non-T cells, and streptavidin nanobeads bind the labeled fraction. A compatible magnetic tube separator retains bead-bound non-target cells against the tube wall. The unlabeled CD3+ T-cell fraction remains in suspension and is poured into a fresh tube without a separation column.
Supplied technical documentation states purity above 90% and up to 99% under tested conditions; one representative flow-cytometry example reached 98.7% post-sort CD3+ purity. These values are representative, not guaranteed. Donor biology, PBMC quality, input composition, handling, separator performance, and collection technique can affect each result.
The documented core separation includes 10 minutes of cocktail labeling, 5 minutes of nanobead binding, and a 3–5 minute magnetic hold. PBMC preparation, cell counting, optional washing, final centrifugation, and downstream quality control are not included.
Keep the cell suspension at or below 1×10⁸ cells/mL and scale reagents proportionally. For 1×10⁷ cells in 100 µL, the documented starting point is 10 µL isolation cocktail and 10 µL nanobeads. When processing fewer than 1×10⁷ cells, use the 1×10⁷-cell reagent amounts unless a laboratory-validated adjustment has been established.
Use the specified FACS buffer: 1× PBS with 2% FBS and 0.02% EDTA. Cocktail labeling and nanobead binding are performed at room temperature. Mix the nanobead suspension for 5–30 seconds before use and remix midway through the 5-minute binding step if settling is observed.
A wash is optional for some patient-derived PBMC samples and may be omitted for typical healthy-donor PBMCs according to the supplied documentation. When used, add five volumes of buffer, centrifuge at 400×g for 5 minutes, discard the supernatant, and restore the original suspension volume before adding nanobeads.
Before separation, bring a 5 mL-tube sample to 2.5 mL or a 15 mL-tube sample to 7.5 mL. Hold for 3 minutes in a compatible single-position magnet or 5 minutes in a compatible multi-position magnet. Use the tube and separator combination specified for the selected format.
While the tube remains in the magnet, tilt the separator and pour the suspended fraction into a fresh collection tube without disturbing retained cells. For culture or analysis, an optional 400×g, 10-minute centrifugation can be used. Verify CD3+ purity, viable-cell recovery, and total yield with a laboratory-validated flow-cytometry panel.
Check PBMC viability and aggregates, input concentration, complete reagent mixing, room-temperature incubation timing, nanobead settling, separator and tube compatibility, final sample volume, magnetic hold time, and pour-off technique. Excess input or disturbing the retained fraction can reduce purity; over-retention or incomplete transfer can reduce recovery.

Related topics covered:

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

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