Mouse CD4+ T Cell Sorting Kit - CS-0030
CS-0030

Mouse CD4+ T Cell Sorting Kit

Column-Based Negative Mouse

The Cell Sorts CS-0030 Mouse CD4 T Cell Isolation Kit uses column-assisted negative selection to isolate untouched CD4⁺ T cells from mouse spleen with 90–99% purity in ~20 minutes. Add 10 μL Mouse CD4 T Cell Isolation Cocktail to label non-target cells at room temperature for 10 min, then 10× FACS Buffer wash and 500g centrifugation for 5 min. Add 10 μL Ultra-Nanobeads for 5 min at room temperature to bind labeled non-target cells.

Load onto an L separation column equilibrated with 3 mL FACS Buffer and collect CD4⁺ T cells as they drip through by gravity. Wash with another 3 mL buffer for maximum yield. No bead contact with target cells — isolated cells are immediately ready for Th1/Th2/Th17 polarization, cytokine profiling, autoimmune disease models, and infectious disease research.

Sorting Purity
90–99%
Sorting Time
25–35 min

Product Features of the Mouse CD4+ T Cell Sorting Kit

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

Column-assisted separation — Magnetic column enables efficient depletion of non-target cells with high recovery rate
No bead residue on target cells — Isolated CD4+ T cells are bead-free and ready for immediate downstream use
Purity 90–99% — High-efficiency negative selection for ultra-pure CD4+ T cell populations
High recovery rate — Optimized column format maximizes yield of viable CD4+ T cells
Suitable for large samples — Column-based format handles larger cell numbers with consistent performance
Native state preservation — Antibodies and beads do not contact CD4+ T cells, maintaining their original functional state

The Mouse CD4+ T Cell Sorting Kit uses column-based negative selection to isolate untouched Target Cells from human PBMCs. To achieve these results, follow this simple 4-step protocol that takes just 25–35 min from start to finish. No centrifugation and no lengthy incubations. Furthermore, this streamlined protocol ensures consistent results across experiments.

How the Mouse CD4+ T Cell Sorting Kit Works

Simple 4-step protocol for rapid cell isolation

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Step 1: Sample Preparation - Prepare single-cell suspension for magnetic cell sorting

Step 1: Sample Preparation

Grind mouse spleen through a 70 μm sterile nylon mesh filter to prepare a single-cell suspension. Count cells and resuspend 1×10⁷ cells in 100 μL FACS Buffer (scale proportionally for higher cell numbers).

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Step 2: Antibody Cocktail Incubation - Label non-target cells with biotin-conjugated antibodies

Step 2: Antibody Labeling

Add 10 μL Mouse CD4 T Cell Isolation Cocktail to the sample, mix gently by pipetting 2–3 times, and incubate at room temperature for 10 min to label non-CD4⁺ T cells.

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Step 3: Magnetic Nanobead Binding - Streptavidin nanobeads bind to labeled non-target cells

Step 3: Magnetic Bead Binding & Wash

Wash cells with 10× volume of FACS Buffer, centrifuge at 500g for 5 min, and resuspend in 100 μL. Vortex Ultra-Nanobeads for 5–30 s, add 10 μL to the sample, mix gently, and incubate at room temperature for 5 min to bind labeled non-target cells.

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Step 4: Column Separation - Purified target cells collected via column or magnetic separator

Step 4: Column-Based Separation & Collection

Place an L separation column into the magnetic field and equilibrate with 3 mL FACS Buffer. Load the cell suspension onto the column and collect CD4⁺ T cells as they drip through by gravity. Add another 3 mL FACS Buffer to wash the column and collect residual target cells for maximum yield.

Applications for the Mouse CD4+ T Cell Sorting Kit

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

Mouse Immunology Research — CD4+ T helper cell activation, proliferation, and cytokine production studies Autoimmune Disease Models — EAE, colitis, and other mouse autoimmune model CD4+ T cell research Infectious Disease Research — Virus-specific CD4+ T cell response analysis in mouse infection models Cancer Immunology — Tumor microenvironment CD4+ T cell studies and immune response evaluation Flow Cytometry — High-purity CD4+ T cell populations ready for staining and analysis Cell Culture — Bead-free, unactivated CD4+ T cells ideal for in vitro expansion and manipulation Drug Discovery — CD4+ T cell-targeted drug screening in syngeneic and transgenic mouse models

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

  • Mouse CD4 T Cell Isolation Cocktail
  • Ultra-Nanobeads
  • User manual

Storage & Shelf Life

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

Mouse CD4+ T Cell Sorting Kit — Frequently Asked Questions

Everything you need to know about the CS-0030 Mouse CD4+ T Cell Sorting Kit

Click each question to expand the answer

The CS-0030 Mouse CD4 T Cell Isolation Kit uses column-assisted negative selection. A biotin-conjugated antibody cocktail labels non-target cells (CD8+ T cells, B cells, NK cells, monocytes, granulocytes) in mouse spleen single-cell suspensions. Ultra-Nanobeads bind the labeled cells, and the mixture is applied to an L separation column under a magnetic field. Non-CD4+ T cells are retained in the column while untouched CD4+ T cells drip through by gravity.
Purity of isolated CD4+ T cells typically reaches 90–99%, as validated by flow cytometry using CD4-FITC (Clone GK1.5) and CD3-PE antibodies. The optimized antibody cocktail efficiently depletes all non-CD4+ cell populations while maximizing recovery of viable, functional CD4+ T cells from mouse spleen.
The entire column-assisted protocol takes approximately ~20 minutes from start to finish: 10 min for antibody cocktail incubation, 10 min for nanobead binding and wash, and ~10 min for column separation.
Antibodies and Ultra-Nanobeads bind exclusively to non-target cells—they never touch CD4+ T cells. This untouched approach ensures full preservation of the T cell receptor, CD4 co-receptor, and native functional states. The column format provides gentle handling with high recovery, making isolated cells ideal for downstream functional assays.
Step 1: Grind mouse spleen through a 70 μm nylon mesh filter. Resuspend 1×10⁷ cells in 100 μL FACS Buffer.
Step 2: Add 10 μL Mouse CD4 T Cell Isolation Cocktail, mix, incubate at room temperature for 10 min.
Step 3: Wash with 10× volume FACS Buffer, centrifuge at 500g for 5 min, resuspend in 100 μL. Add 10 μL Ultra-Nanobeads, mix, incubate at room temperature for 5 min.
Step 4: Place an L column in the magnetic field, equilibrate with 3 mL FACS Buffer. Load suspension, collect CD4+ T cells as they drip through. Wash with 3 mL buffer for maximum yield.
Total time: ~20 minutes.
Yes, the CS-0030 kit is ideal for Th polarization research. The untouched, bead-free CD4+ T cells maintain full TCR and CD4 signaling functionality, ensuring optimal conditions for polarizing into Th1, Th2, or Th17 subsets under specific cytokine conditions.
The antibody cocktail targets CD8+ T cells, B cells, NK cells, monocytes, and granulocytes for depletion while leaving CD4+ T cells completely untouched. This comprehensive depletion of non-CD4+ populations ensures high purity and recovery.
Isolated CD4+ T cells are immediately ready for: Th1/Th2/Th17 polarization, cytokine profiling, autoimmune disease models, infectious disease research, flow cytometry, cell culture, and functional assays. The untouched, bead-free state means no additional washing steps are required.
We recommend flow cytometry using CD4-FITC (Clone GK1.5), CD3-PE, and CD8a-PerCP-Cy5.5 antibodies. Typical results show 90–99% CD4+ cells. Viability can be assessed using Trypan blue exclusion or 7-AAD staining. For best results, gate on lymphocytes by forward and side scatter.
The CS-0030 uses column-assisted negative selection (~20 min, 90–99% purity) with a wash/centrifuge step and gravity drip collection, ideal for higher consistency. The CS-0029 uses column-free negative selection (18–20 min, up to 99% purity) with pour-off collection, offering faster processing with no columns. Both yield untouched, bead-free CD4+ T cells.

Related topics covered:

column-based negative selection Target Cell isolation Target Cell purity verification Target Cell sorting protocol column-based vs column-free single-cell analysis (scRNA-seq) Target Cell isolation markers PBMC Target Cell sorting

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