CS-0048

Mouse CD3ε NanoBeads Sorting Kit

Column-Based Positive Mouse

The Cell Sorts Mouse CD3ε NanoBeads Sorting Kit (CS-0048) uses column-based positive selection to isolate total T lymphocytes expressing the CD3ε subunit from mouse splenocytes or peripheral blood single-cell suspensions with 95–99% purity in 20–30 min. Anti-mouse NanoBeads conjugated to the T cell marker bind target populations, which are retained in a magnetic separation column; unlabeled non-target cells (B lymphocytes, NK cells, monocytes, granulocytes) flow through. Eluted T cells show >90% recovery and >98% viability — ready for T cell activation assays, CAR-T development, cytokine profiling, and TCR repertoire sequencing.

Compared to density gradient centrifugation (40–60% purity) or FACS (expensive, low throughput), this Cell Sorts kit delivers 95–99% purity with >90% recovery. The releasable NanoBeads leave cells bead-free in 10 min — ideal for single-cell RNA sequencing (scRNA-seq), electroporation, and CRISPR gene editing. Captures both CD4⁺ helper and CD8⁺ cytotoxic T cells for comprehensive mouse T cell research.

Sorting Purity

≥95%

Sorting Time

~30 min

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Product Features of the Mouse CD3ε NanoBeads Sorting Kit

The Mouse CD3ε NanoBeads Sorting Kit delivers industry-leading performance for rapid CD3ε+ T Cell isolation from PBMCs via column-based positive selection.

Column-assisted positive selection — The pan-T cell marker antibody-conjugated NanoBeads directly label T lymphocytes for efficient magnetic capture in a separation column from mouse splenocytes or blood

Purity 95–99% — High-efficiency positive selection delivers ultra-pure T lymphocyte populations from mouse single-cell suspensions, validated by flow cytometry

Releasable from column — Elute captured T cells by plunger press; optional Release Buffer removes NanoBeads for bead-free downstream processing

Scalable format — Small column handles 1×10⁷ cells; medium column accommodates up to 1×10⁹ total cells. From bench immunology to large-scale adoptive cell therapy studies

Multi-sample compatible — Optimized for mouse peripheral blood and splenic lymphocyte single-cell suspensions. Compatible with C57BL/6, BALB/c, and common mouse strains

Flow cytometry validated — Compatible with CD3-APC (Clone 17A2), CD4-FITC, and CD8-APC staining for immediate purity verification and CD4/CD8 subset characterization

The Mouse CD3ε NanoBeads Sorting Kit uses column-based positive selection to isolate purified CD3ε+ T Cells from human PBMCs. To achieve these results, follow this simple 4-step protocol that takes just ~30 min from start to finish. No centrifugation and no lengthy incubations. Furthermore, this streamlined protocol ensures consistent results across experiments.

How the Mouse CD3ε NanoBeads Sorting Kit Works

Simple 4-step protocol for rapid cell isolation

Step 1: Sample Preparation

Prepare single-cell suspension from mouse peripheral blood or lymphoid organs (spleen). Count cells, resuspend 1×10⁷ cells in 100 μL cold separation buffer (2–8°C).

Step 2: Biotin Antibody Labeling & Magnetic Bead Binding

Add 1.5 μL Biotin Anti-Mouse CD3ε Antibody, mix, incubate 4°C 10 min. Wash, resuspend 90 μL. Add 10 μL Anti-Biotin NanoBeads, mix, incubate 4°C 15 min. Wash, resuspend 500 μL.

Step 3: Column-Based Magnetic Separation

Place pre-washed column in magnetic separator. Load cell suspension, allow flow-through. Wash column 3× with separation buffer to remove unlabeled non-target cells. Magnetically labeled T cells retained.

Step 4: Target Cell Elution & Collection

Remove column from magnet, place over collection tube, add separation buffer (1 mL small / 5 mL medium), flush captured T cells with plunger. Ready for T cell activation, CAR-T, cytokine profiling, or scRNA-seq.

Applications for the Mouse CD3ε NanoBeads Sorting Kit

This Mouse CD3ε NanoBeads Sorting Kit supports a wide range of downstream applications for CD3ε+ T Cell isolation and column-based positive selection workflows from mouse splenocytes or lymph node cells.

Mouse Immunology Research — T cell activation, proliferation, and cytokine production studies Cell Therapy R&D — CAR-T and TCR-T engineering from mouse CD3ε+ T cells Autoimmune Disease Studies — EAE, colitis, and other mouse autoimmune model T cell research Infectious Disease Research — Virus-specific T cell response analysis in mouse infection models Flow Cytometry — High-purity CD3ε+ T cell populations ready for CD3-APC staining and analysis Cell Culture — Bead-bound CD3ε+ T cells for activation, expansion, and functional manipulation Drug Discovery — T cell-targeted drug screening in syngeneic and transgenic mouse models

In addition, the Mouse CD3ε NanoBeads Sorting Kit is compatible with a wide range of downstream applications, making it a versatile tool for your immunology research workflow.

Kit Components

Each Mouse CD3ε NanoBeads Sorting Kit contains everything needed for high-purity column-based positive selection from human PBMCs.

Biotin Anti-Mouse CD3ε Antibody
Anti-Biotin Nano Beads
User manual

Storage & Shelf Life

Storage

Store at 2-8°C protected from light

Shelf Life

12 months

Mouse CD3ε NanoBeads Sorting Kit — Frequently Asked Questions

Everything you need to know about the CS-0048 Mouse CD3ε NanoBeads Sorting Kit

Click each question to expand the answer

The CS-0048 Mouse CD3ε NanoBeads Sorting Kit from Cell Sorts uses column-based positive selection. Anti-mouse CD3ε antibody-conjugated NanoBeads directly bind to the T cell surface marker. The cell-bead mixture is applied to a magnetic separation column, where T lymphocytes are retained. Unlabeled non-target cells flow through. The column is then removed from the magnet, and purified T cells are eluted by plunger press.

Purity of isolated mouse T cells typically reaches 95–99%, as validated by flow cytometry using anti-mouse CD3 antibody staining. The NanoBeads positive selection protocol efficiently enriches total T cells from mouse splenocytes and peripheral blood while depleting B cells, NK cells, and myeloid cells.

The CS-0048 kit is optimized for mouse splenocytes and peripheral blood lymphocyte suspensions. Prepare a single-cell suspension by gentle mechanical disruption through a 70 μm cell strainer. Perform red blood cell lysis if needed (e.g., ACK lysis buffer). The kit is compatible with C57BL/6, BALB/c, and other common mouse strains used in immunology research.

Step 1: Prepare a single-cell suspension from mouse blood or spleen.
Step 2: Count cells and resuspend at recommended density.
Step 3: Incubate with Biotin Anti-Mouse CD3ε Antibody, then Anti-Biotin NanoBeads at 4°C.
Step 4: Load onto pre-washed magnetic column.
Step 5: T cells are retained; unlabeled cells flow through.
Step 6: Elute purified T cells by plunger press.
Total time: 20–30 minutes.

Isolated mouse T cells are immediately ready for: T cell activation and proliferation assays (anti-CD3/CD28 stimulation), cytokine profiling (IFN-γ, IL-2, TNF-α), tumor immunology studies (TIL analysis, tumor killing assays), CAR-T cell therapy development in mouse models, adoptive T cell transfer experiments, CRISPR gene editing, single-cell RNA sequencing (scRNA-seq), TCR repertoire sequencing, and flow cytometry analysis.

Yes. The CS-0048 kit uses anti-CD3 targeting, which labels all T cells expressing the T cell receptor complex. This means both CD4+ helper T cells and CD8+ cytotoxic T cells are captured. After isolation, you can further separate CD4+ and CD8+ subsets using secondary sorting or analyze the ratio by flow cytometry. For mouse CD4+ or CD8+ specific isolation, consider complementary Cell Sorts sorting strategies.

The CS-0048 kit uses nano-sized magnetic beads (NanoBeads) that offer higher binding efficiency and lower non-specific binding compared to conventional micron-sized beads. Their small size ensures minimal interference with the T cell receptor complex and signaling pathways, preserving full T cell functionality—critical for activation, cytotoxicity, and cytokine production assays.

We recommend flow cytometry using anti-mouse CD3-APC (Clone 17A2), CD4-FITC, CD8-APC, and B220-PerCP-Cy5.5 antibodies. Typical results show >95% CD3+ cells with minimal B220+ B cell contamination. Additional markers (CD44, CD62L for effector/memory distinction, FoxP3 for Treg, PD-1 for exhaustion) can be used for subset characterization. Viability can be assessed using Trypan blue or 7-AAD staining.

Absolutely. Since isolated mouse T cells are bead-free after NanoBeads release, they are immediately compatible with single-cell RNA sequencing (scRNA-seq), TCR repertoire sequencing, and other single-cell technologies. This enables high-resolution profiling of T cell heterogeneity and immune repertoire analysis at single-cell resolution in mouse models. Cell Sorts protocols are validated for 10x Genomics and other major scRNA-seq platforms.

The CS-0048 kit targets mouse T cells using positive selection with column-based capture, designed for mouse splenocytes. The CS-0004 kit targets human CD3+ T cells using negative selection (untouched approach), designed for human PBMCs. CS-0048 uses direct NanoBeads labeling of T cells; CS-0004 uses antibody cocktail + streptavidin nanobeads to deplete non-T cells. Both achieve 90-99% purity.