CellSorts CS-0038 Mouse Naïve CD4+ T Cell Sorting Kit — column-free negative selection with FcR blocking, no RBC lysis, up to 99% purity, 18-20 min
CS-0038

Mouse Naïve CD4+ T Cell Sorting Kit

Column-Free Negative Mouse

The Cell Sorts CS-0038 Mouse Naïve CD4+ T Cell Sorting Kit uses column-free negative selection with FcR blocking to isolate untouched naïve CD4+ T cells from mouse spleen with up to 99% purity in 18–20 min. No RBC lysis required. Mouse CD16/CD32 Blocker plus Mouse CD4 T Cell Isolation Cocktail and Mouse Memory T Cell Depletion Cocktail (100 μL/mL each) label non-target cells and deplete memory CD4 T cells at RT for 10 min. Streptavidin Nanobeads (100 μL/mL) bind for 5 min; magnetic separator 3–5 min, then pour off purified naïve CD4+ T cells.

No columns, no bead or antibody contact with target cells. FcR blocking plus dual antibody labeling ensures high purity of the naïve subset (CD62L+CD44−). Isolated naïve CD4+ T cells are untouched — immediately ready for T cell differentiation, Th polarization, primary immune response studies, vaccine development, flow cytometry, scRNA-seq, and adoptive cell therapy.

Sorting Purity
Up to 99%
Sorting Time
18–20 min

Product Features of the Mouse Naïve CD4+ T Cell Sorting Kit

The Mouse Naïve CD4+ T Cell Sorting Kit delivers industry-leading performance for rapid Naïve CD4+ T Cell isolation from PBMCs via column-free negative selection.

Column-free separation — No column needed, streamlined workflow; simply pour target cells into a new tube
No RBC lysis required — Optimized protocol eliminates red blood cell lysis step, saving time and preserving cell viability
Zero contact with target cells — Antibodies and beads bind non-naïve CD4 T cells only, preserving naïve CD4+ T cells in their native, unactivated state
Purity up to 99% — High-efficiency negative selection for ultra-pure naïve CD4+ T cell populations
18–20 min fast sorting — Rapid protocol from mouse spleen to purified naïve CD4+ T cells
Easy handling — Minimal steps, user-friendly procedure suitable for any lab
Ready for downstream use — Isolated cells are immediately compatible with flow cytometry, cell culture, and functional assays

The Mouse Naïve CD4+ T Cell Sorting Kit uses column-free negative selection to isolate untouched Naïve CD4+ T Cells from human PBMCs. To achieve these results, follow this simple 4-step protocol that takes just 18–20 min from start to finish. No centrifugation and no lengthy incubations. Furthermore, this streamlined protocol ensures consistent results across experiments.

How the Mouse Naïve CD4+ T Cell Sorting Kit Works

Simple 4-step protocol for rapid cell isolation

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Step 1 of naïve CD4+ T cells sorting: Sample Preparation - Prepare single-cell suspension from mouse tissue for CS-0038 Mouse Naïve CD4+ T Cell Sorting Kit

Step 1: Sample Preparation

Harvest mouse spleen, grind through a 70 μm sterile nylon mesh filter. Count cells and resuspend in FACS Buffer at 1×10⁸ cells/mL (e.g., 1×10⁷ cells in 100 μL). No RBC lysis required.

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Step 2 of naïve CD4+ T cells sorting: Antibody Cocktail Labeling - Biotin-conjugated antibodies target non-naïve CD4+ cells for depletion with CS-0038

Step 2: FcR Blocking & Dual Antibody Labeling

Add Mouse CD16/CD32 Blocker (100 μL/mL) to block Fc receptors. Then add Mouse CD4 T Cell Isolation Cocktail (100 μL/mL) and Mouse Memory T Cell Depletion Cocktail (100 μL/mL), mix gently by pipetting 2–3 times, incubate RT 10 min.

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Step 3 of naïve CD4+ T cells sorting: Magnetic Nanobead Binding - Streptavidin nanobeads bind labeled non-target cells for magnetic removal with CS-0038

Step 3: Magnetic Bead Binding

Vortex Streptavidin Nanobeads 5–30 s, add 100 μL per mL of cell suspension, mix gently, incubate RT 5 min to bind all labeled non-target cells.

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Step 4 of naïve CD4+ T cells sorting: Column-Free Separation - Purified naïve CD4+ T cells collected by magnetic separator pour-off for CS-0038

Step 4: Column-Free Separation & Collection

Dilute with FACS Buffer to 2.5 mL (5 mL tube) or 7.5 mL (15 mL tube). Place on magnetic separator 3–5 min, then pour off untouched purified naïve CD4⁺ T cells into a fresh collection tube.

Applications for the Mouse Naïve CD4+ T Cell Sorting Kit

This Mouse Naïve CD4+ T Cell Sorting Kit supports a wide range of downstream applications for Naïve CD4+ T Cell isolation and column-free negative selection workflows from mouse splenocytes or lymph node cells.

NK Cell Biology Research — Cytotoxicity, ADCC, cytokine production, and receptor expression studies in mouse models Cancer Immunology — Tumor-NK cell interaction studies, immune surveillance research Infectious Disease Research — Viral pathogen-NK cell response analysis in mouse infection models Immunotherapy R&D — CAR-NK and adoptive NK cell therapy development Flow Cytometry — High-purity NK cell populations ready for CD49b/NK1.1 staining and analysis Cell Culture — Unactivated, bead-free NK cells ideal for in vitro expansion and manipulation Drug Discovery — NK cell-mediated cytotoxicity assays, immunomodulatory drug screening

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

  • Mouse CD16/CD32 Blocker
  • Mouse CD4 T Cell Isolation Cocktail
  • Mouse Memory T Cell Depletion Cocktail
  • Streptavidin Nanobeads
  • User manual

Storage & Shelf Life

Storage
Store at 2-8°C protected from light
Shelf Life
12 months

Mouse Naïve CD4+ T Cell Sorting Kit — Frequently Asked Questions

Everything you need to know about the CS-0038 Mouse Naïve CD4+ T Cell Sorting Kit

Click each question to expand the answer

The CS-0038 Mouse Naïve CD4+ T Cell Sorting Kit from Cell Sorts uses column-free negative selection. Non-target cells (CD8+ T cells, B cells, NK cells, monocytes, granulocytes, erythrocytes, memory/effector T cells, Tregs) are labeled with a biotin-conjugated antibody cocktail and removed by magnetic separation, while untouched naïve CD4+ T cells remain in the supernatant. This ensures no beads or antibodies ever contact the target cells, preserving their native functional state.
Purity of isolated naïve CD4+ T cells typically reaches up to 99%, as validated by flow cytometry using CD4-FITC (Clone GK1.5), CD62L-PE, CD44-APC, CD45RA staining. The optimized negative selection protocol with FcR blocking (Mouse CD16/CD32 Blocker) and dual antibody labeling (Mouse CD4 T Cell Isolation Cocktail + Mouse Memory T Cell Depletion Cocktail) efficiently depletes all non-target cell populations while maximizing recovery of viable, functional naïve CD4+ T cells from mouse spleen. No RBC lysis is required, preserving cell viability.
The CS-0038 kit is optimized for mouse spleen and lymph node cell suspensions. Prepare a single-cell suspension by gentle mechanical disruption through a 70 μm sterile nylon mesh filter. No red blood cell lysis is required — the optimized protocol eliminates the ACK lysis step, saving time and preserving cell viability. 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 sterile nylon mesh filter. Count cells and resuspend in FACS Buffer at 1×10⁸ cells/mL (e.g., 1×10⁷ cells in 100 μL). No RBC lysis required.
Step 2: FcR Blocking & Dual Antibody Labeling. Add Mouse CD16/CD32 Blocker (100 μL/mL) to block Fc receptors. Then add Mouse CD4 T Cell Isolation Cocktail (100 μL/mL) and Mouse Memory T Cell Depletion Cocktail (100 μL/mL), mix gently by pipetting 2–3 times, incubate at room temperature for 10 min.
Step 3: Magnetic Bead Binding. Vortex Streptavidin Nanobeads for 5–30 s, add 100 μL per mL of cell suspension, mix gently, incubate at room temperature for 5 min.
Step 4: Column-Free Separation & Collection. Dilute with FACS Buffer to 2.5 mL (5 mL tube) or 7.5 mL (15 mL tube). Place on magnetic separator for 3–5 min, then pour off the untouched, purified naïve CD4⁺ T cells into a fresh collection tube.
Total time: 18–20 min.
Isolated naïve CD4+ T cells are immediately ready for: T cell differentiation studies, Th polarization from naïve precursors, primary immune response, 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 CD4+ 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 CD8a, CD19, CD11b, CD49b, Ly-6G, Ter-119, CD44, CD25 on non-target cells—effectively labeling CD8+ T cells, B cells, NK cells, monocytes, granulocytes, erythrocytes, memory/effector T cells, Tregs for depletion while leaving naïve CD4+ T cells completely untouched. This comprehensive marker panel ensures high purity and recovery of the target cell population.
We recommend flow cytometry using CD4-FITC (Clone GK1.5), CD62L-PE, CD44-APC, CD45RA antibodies. Typical results show up to 99% naïve CD4⁺ T cells (CD62L⁺CD44⁻) 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 CD4+ 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-0038 uses column-free negative selection (18–20 min) with FcR blocking (Mouse CD16/CD32 Blocker) and dual antibody labeling (Mouse CD4 T Cell Isolation Cocktail + Mouse Memory T Cell Depletion Cocktail), achieving up to 99% purity with no RBC lysis required. Column-based kits (25–35 min) offer higher consistency for larger sample sizes. Both yield untouched, bead-free naïve CD4⁺ T cells. Choose CS-0038 for rapid processing with FcR-blocked, memory-depleted, high-purity output; choose column-based for larger-scale isolation.

Related topics covered:

column-free negative selection Naïve CD4+ T Cell isolation Naïve CD4+ T Cell purity verification Naïve CD4+ T Cell sorting protocol column-free isolation single-cell analysis (scRNA-seq) Naïve CD4+ T Cell isolation markers PBMC Naïve CD4+ T Cell sorting

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