CS-0005

Human CD4+ T Cell Sorting Kit

Column-Free Negative Human

Cell Sorts™ Human CD4+ T Cell Sorting Kit (CS-0005) enriches untouched CD4+ T cells from human PBMCs using column-free negative selection. A biotin-conjugated cocktail labels non-target cells, streptavidin nanobeads bind those cells, and a tube magnet retains them. The unlabeled CD4+ T-cell fraction is recovered from the suspension without direct bead or antibody labeling.

Supplied technical documentation reports purity above 90%, with results up to 98%; one representative test enriched CD3+CD4+ cells from 21.6% to 95.6%. The documented core workflow is 18–20 minutes after PBMC preparation and counting. Recovered cells may support flow cytometry, cell culture, activation, cytokine, polarization, and single-cell studies after laboratory-specific purity, viability, recovery, and yield checks. For research use only.

Sorting Purity

Above 90%, up to 98%

Sorting Time

18–20 min

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Product Features of the Human CD4+ T Cell Sorting Kit

CS-0005 uses a documented column-free negative-selection workflow to enrich untouched human CD4+ T cells from density-gradient-prepared PBMCs.

Column-free negative selection — Non-target cells are magnetically retained in a compatible tube separator; no separation column is required

Untouched CD4+ T-cell fraction — Target cells are not directly labeled with the depletion cocktail or nanobeads

Documented purity above 90%, up to 98% — A representative PBMC test enriched CD3+CD4+ cells from 21.6% to 95.6%

18–20 minute core workflow — Includes 10-minute cocktail labeling, 5-minute nanobead binding, magnetic separation, and collection

Defined reagent ratio — 100 µL isolation cocktail and 100 µL nanobeads per mL of cells at up to 1×10⁸ cells/mL

Flexible tube formats — Compatible workflows are documented for 5 mL, 15 mL, and 50 mL tubes with matching magnetic separators

CS-0005 uses column-free negative selection to enrich untouched human CD4+ T cells from PBMCs. The documented core workflow takes approximately 18–20 minutes after PBMC preparation and counting, with purity reported above 90% and up to 98%.

How the Human CD4+ T Cell Sorting Kit Works

Simple 4-step protocol for rapid cell isolation

Step 1: Prepare the PBMC Suspension

Prepare PBMCs by density-gradient centrifugation, count viable cells, and resuspend at up to 1×10⁸ cells/mL. At least 1×10⁷ cells per separation is recommended.

Step 2: Label Non-Target Cells

Add 100 µL of isolation cocktail per mL of cells, mix gently, and incubate for 10 minutes at room temperature. An optional 400×g wash may be appropriate for selected patient-derived PBMC samples.

Step 3: Bind Magnetic Nanobeads

Add 100 µL of nanobeads per mL of cells, mix, and incubate for 5 minutes at room temperature. Add at least five volumes of FACS buffer before magnetic separation.

Step 4: Separate and Recover CD4+ T Cells

Magnetize for 3 minutes with a compatible single-position separator or 5 minutes with a multi-position separator, then transfer the enriched suspension without disturbing retained cells.

Applications for the Human CD4+ T Cell Sorting Kit

This Human CD4+ T Cell Sorting Kit supports a wide range of downstream applications for CD4+ T Cell isolation and column-free negative selection workflows from human PBMCs.

CD4+ T Cell Phenotyping — Prepare enriched populations for multiparameter flow-cytometry panels after purity and viability assessment Activation and Proliferation Studies — Evaluate receptor-driven responses in untouched CD4+ T-cell preparations Helper T Cell Differentiation — Support laboratory-validated Th1, Th2, Th17, or regulatory T-cell polarization workflows Cytokine Analysis — Study cytokine secretion and signaling after appropriate stimulation and controls Cell Culture — Establish downstream expansion or functional-assay workflows using the recovered supernatant fraction Immunology and Host-Response Research — Investigate CD4+ T-cell biology in infectious-disease, inflammatory, or immune-regulation models Single-Cell and Molecular Analysis — Prepare samples for RNA, TCR, or single-cell workflows when input-quality requirements are met Drug Discovery — Assess candidate immunomodulators in laboratory-validated CD4+ T-cell assays

In addition, the Human CD4+ 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 CD4+ T Cell Sorting Kit contains everything needed for high-purity column-free negative selection from human PBMCs.

Human CD4+ T Cell Isolation Cocktail
Streptavidin Nanobeads

Storage & Shelf Life

Storage

Store at 4°C; do not freeze

Shelf Life

Refer to the product label

Human CD4+ T Cell Sorting Kit — Frequently Asked Questions

Everything you need to know about the CS-0005 Human CD4+ T Cell Sorting Kit

Click each question to expand the answer

CS-0005 is designed to enrich untouched human CD4+ T cells from PBMC preparations. The supplied protocol recommends preparing PBMCs by density-gradient centrifugation rather than red-blood-cell lysis. Starting-cell viability, aggregation, donor variation, and the initial CD4+ T-cell frequency can affect performance.

A biotin-conjugated isolation cocktail labels non-target cells, and streptavidin nanobeads bind the labeled population. A compatible tube magnet retains those cells against the tube wall. The unlabeled CD4+ T-cell fraction remains in suspension and is transferred to a fresh tube without a separation column.

The supplied technical documentation reports purity above 90%, with results up to 98%. In one representative PBMC example, the CD3+CD4+ population increased from 21.6% before isolation to 95.6% afterward. These observations are not guaranteed specifications; verify purity, viability, recovery, and yield in each laboratory.

The documented core isolation workflow is approximately 18–20 minutes after PBMC preparation and counting. It includes a 10-minute room-temperature cocktail incubation, a 5-minute nanobead incubation, magnetic separation for the specified tube-and-magnet format, and collection.

For PBMCs prepared at up to 1×10⁸ cells/mL, the documentation specifies 100 µL of isolation cocktail and 100 µL of nanobeads per mL of cell suspension. For 1×10⁷ cells in 100 µL, use 10 µL of each reagent. Scale volumes proportionally and consult the current instructions.

Prepare a uniform suspension at no more than 1×10⁸ cells/mL. The documentation recommends processing at least 1×10⁷ cells per separation; lower inputs should use the reagent volumes specified for 1×10⁷ cells. Available kit formats support total inputs up to 1×10⁸, 5×10⁸, or 1×10⁹ cells.

The documentation describes an optional wash for selected patient-derived PBMC samples: add five volumes of FACS buffer, centrifuge at 400×g for 5 minutes, remove the supernatant, and restore the original volume. Healthy-donor PBMCs may proceed without this step. Validate the decision for the sample type and study design.

The protocol provides workflows for 5 mL, 15 mL, and 50 mL tubes with compatible single-position or multi-position magnetic separators. The documented magnetic hold is 3 minutes for a single-position separator and 5 minutes for a multi-position separator. Confirm the tube volume and separator combination before use.

Store the isolation cocktail and streptavidin nanobeads at 4°C and do not freeze. The supplied documentation also specifies shipment at 4°C. Follow the current product label for lot-specific shelf life and handling instructions.

After confirming purity and viability, the enriched fraction may support flow cytometry, cell culture, activation and proliferation assays, cytokine analysis, helper-T-cell polarization, TCR or molecular studies, and single-cell workflows. Each downstream application requires laboratory-specific qualification.

Review PBMC viability and aggregation, input concentration, cocktail and bead mixing, room-temperature incubation times, buffer quality, tube-to-magnet compatibility, magnetic hold time, and transfer technique. Excess input, incomplete resuspension, or disturbing retained cells can reduce purity or viable-cell recovery.