CS-0047

Mouse Naïve T Cell Sorting Kit

Column-Free Negative Mouse

The Cell Sorts CS-0047 Mouse Naïve T Cell Sorting Kit uses column-free negative selection to isolate untouched naïve CD3+ T cells from mouse spleen and lymph node with highly pure populations in ~20–25 min. Mouse Pan-Naïve T Cell Isolation Cocktail (3 μL) labels non-target cells at RT for 5 min, followed by Mouse Pan-Naïve T Beads Streptavidin 1.0-N (10 μL) for 5 min. Magnetic separator 5 min; repeat once and pool both fractions for higher recovery.

No columns, no bead or antibody contact with target cells. Multi-tissue compatible — validated on mouse spleen and lymph node samples. Isolated naïve T cells are untouched in their unprimed, quiescent state — immediately ready for T cell development, primary immune response, vaccine development, flow cytometry, scRNA-seq, and adoptive cell therapy.

Sorting Purity

Up to 95.63%

Sorting Time

20-25 min

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Product Features of the Mouse Naïve T Cell Sorting Kit

The Mouse Naïve T Cell Sorting Kit delivers industry-leading performance for rapid Naïve T Cell isolation from PBMCs via column-free negative selection.

Negative selection — No antibody or bead contact with naïve T cells, preserving their unprimed, quiescent state

High purity — Rapid and simple protocol delivers highly pure naïve CD3+ T cell populations

Multi-tissue compatible — Validated on mouse spleen and lymph node samples

Ready for downstream use — Isolated cells are immediately compatible with flow cytometry, cell culture, and functional assays

Convenient assay format — Pre-optimized reagent volumes per assay streamline workflow planning

The Mouse Naïve T Cell Sorting Kit uses column-free negative selection to isolate untouched Naïve T Cells from human PBMCs. To achieve these results, follow this simple 4-step protocol that takes just 20-25 min from start to finish. No centrifugation and no lengthy incubations. Furthermore, this streamlined protocol ensures consistent results across experiments.

How the Mouse Naïve T Cell Sorting Kit Works

Simple 4-step protocol for rapid cell isolation

Step 1: Sample Preparation

Harvest mouse spleen, grind through a 70 μm strainer, wash with pre-cooled PBS, centrifuge 300×g 5 min. Resuspend in separation buffer at 2×10⁸ cells/mL.

Step 2: Antibody Cocktail Labeling

Aliquot 50 μL cell suspension (1×10⁷ cells) into a 2 mL EP tube. Add 3 μL Mouse Pan-Naïve T Cell Isolation Cocktail, mix well, RT 5 min. Wash with separation buffer, centrifuge 300×g 5 min, resuspend in 50 μL separation buffer.

Step 3: Magnetic Bead Binding

Add 10 μL pre-washed Mouse Pan-Naïve T Beads Streptavidin 1.0-N, mix gently by pipetting 6–8 times, RT 5 min to bind non-target cells.

Step 4: Column-Free Separation & Collection

Dilute with separation buffer to 2.5 mL, mix thoroughly by pipetting 6–8 times until no visible bead aggregates remain. Place on magnetic separator for 5 min. Transfer supernatant to clean tube — purified Pan-Naïve T cell fraction. Repeat once and pool both fractions for higher recovery.

Applications for the Mouse Naïve T Cell Sorting Kit

This Mouse Naïve T Cell Sorting Kit supports a wide range of downstream applications for Naïve T Cell isolation and column-free negative selection workflows from mouse splenocytes or lymph node cells.

Naïve T Cell Biology — De novo T cell activation, proliferation, and differentiation studies T Cell Differentiation Studies — Th1, Th2, Th17, Treg, and other lineage polarization from a pure Naïve starting population Cell Therapy R&D — CAR-T and TCR-T engineering from Naïve precursors for enhanced persistence and potency Vaccine Development — De novo immune response modeling and epitope discovery Flow Cytometry — High-purity Naïve T cell populations ready for CD3/CD44/CD62L staining and analysis Cell Culture — Unactivated, bead-free Naïve T cells ideal for in vitro expansion and manipulation Autoimmune Disease Studies — Naïve T cell self-tolerance and dysregulation mechanisms in mouse models

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

Mouse Pan-Naïve T cell negative selection antibody cocktail
Magnetic beads
User manual

Storage & Shelf Life

Storage

Store at 2-8°C protected from light

Shelf Life

12 months

Mouse Naïve T Cell Sorting Kit — Frequently Asked Questions

Everything you need to know about the CS-0047 Mouse Naïve T Cell Sorting Kit

Click each question to expand the answer

The CS-0047 Mouse Naïve T Cell Sorting Kit from Cell Sorts uses column-free negative selection. Mouse Pan-Naïve T Cell Isolation Cocktail (3 μL) labels non-naïve T cells (B cells, NK cells, monocytes, granulocytes, erythrocytes, memory/effector T cells, Tregs) at room temperature for 5 min. After washing and centrifugation (300×g, 5 min), Mouse Pan-Naïve T Beads Streptavidin 1.0-N (10 μL) bind labeled cells for 5 min. A magnetic separator depletes non-targets for 5 min; purified naïve T cells are carefully transferred to a clean tube. Repeat once and pool both fractions for higher recovery — no beads or antibodies ever contact the target cells.

Purity of isolated naïve T cells typically reaches highly pure naïve CD3⁺ T cell populations, as validated by flow cytometry using CD3-APC (Clone 17A2), CD4-FITC, CD8a-PerCP-Cy5.5, CD62L-PE, CD44-APC staining. The optimized negative selection protocol with Mouse Pan-Naïve T Cell Isolation Cocktail and Streptavidin 1.0-N beads efficiently depletes all non-target cell populations while maximizing recovery of viable, functional naïve T cells from mouse spleen and lymph node samples.

The CS-0047 kit is multi-tissue compatible and validated on mouse spleen and lymph node samples. Prepare a single-cell suspension by grinding through a 70 μm strainer, wash with pre-cooled PBS, and centrifuge at 300×g for 5 min. Resuspend in separation buffer at 2×10⁸ cells/mL. The kit is compatible with C57BL/6, BALB/c and other common mouse strains used in immunology research.

Step 1: Sample Preparation. Harvest mouse spleen, grind through a 70 μm strainer, wash with pre-cooled PBS, and centrifuge at 300×g for 5 min. Resuspend in separation buffer at 2×10⁸ cells/mL.
Step 2: Antibody Cocktail Labeling. Aliquot 50 μL cell suspension (1×10⁷ cells) into a 2 mL EP tube. Add 3 μL Mouse Pan-Naïve T Cell Isolation Cocktail, mix well, incubate at room temperature for 5 min. Wash with separation buffer, centrifuge 300×g 5 min, resuspend in 50 μL separation buffer.
Step 3: Magnetic Bead Binding. Add 10 μL pre-washed Mouse Pan-Naïve T Beads Streptavidin 1.0-N, mix gently by pipetting 6–8 times, incubate at room temperature for 5 min.
Step 4: Column-Free Separation & Collection. Dilute with separation buffer to 2.5 mL, mix thoroughly by pipetting 6–8 times until no visible bead aggregates remain. Place on magnetic separator for 5 min. Transfer supernatant to clean tube — purified Pan-Naïve T cell fraction. Repeat once and pool both fractions for higher recovery.
Total time: ~20–25 min.

Isolated naïve T cells are immediately ready for: T cell development, primary immune response, T cell repertoire diversity, vaccine development, flow cytometry analysis, cell culture and expansion, CRISPR gene editing, single-cell RNA sequencing (scRNA-seq), and adoptive cell therapy studies. Zero bead or antibody interference means no additional washing steps are required.

Antibodies and streptavidin nanobeads bind exclusively to non-target cells—they never touch naïve T cells. This untouched approach ensures full preservation of surface receptors, native functional states, and physiological relevance for downstream functional assays. Cell Sorts column-free technology eliminates column-induced mechanical stress, further enhancing cell viability.

The antibody cocktail targets CD19, CD11b, CD49b, Ly-6G, Ter-119, CD44, CD25 on non-target cells—effectively labeling B cells, NK cells, monocytes, granulocytes, erythrocytes, memory/effector T cells, Tregs for depletion while leaving naïve T cells completely untouched. This comprehensive marker panel ensures high purity and recovery of the target cell population.

We recommend flow cytometry using CD3-APC (Clone 17A2), CD4-FITC, CD8a-PerCP-Cy5.5, CD62L-PE, CD44-APC antibodies. Typical results show 90-99% target cells in the collected fraction. Viability can be assessed using Trypan blue exclusion or 7-AAD staining. For best results, gate on lymphocytes by forward and side scatter, then analyze marker expression within the gated population.

Absolutely. Since isolated naïve T cells are completely bead-free and antibody-free, they are immediately compatible with single-cell RNA sequencing (scRNA-seq) and other single-cell technologies without additional washing steps. Cell Sorts protocols are validated for 10x Genomics and other major scRNA-seq platforms.

The CS-0047 uses column-free negative selection (20-25 min), ideal for speed and convenience with smaller samples. Column-based kits (25–35 min) offer higher consistency for larger sample sizes. Both yield untouched, bead-free naïve T cells with 90-99% purity. Choose CS-0047 for rapid processing; choose column-based for larger-scale isolation.