CellSorts CS-0041 Mouse NK Cell Sorting Kit — column-free negative selection, no RBC lysis, up to 95% purity
CS-0041

Mouse NK Cell Sorting Kit

Column-Free Negative Mouse

The Cell Sorts CS-0041 Mouse NK Cell Sorting Kit uses column-free negative selection to isolate untouched NK cells from mouse spleen with up to 95% purity in 18–20 min. No RBC lysis is required — grind the spleen and filter through a 70 μm sterile nylon mesh. NK Cell Isolation Cocktail (100 μL/mL) labels non-NK cells at room temperature for 10 min, followed by Streptavidin Nanobeads (100 μL/mL) at room temperature for 5 min. Magnet 3 min (single-position) or 5 min (multi-position), then pour off purified NK cells.

No columns, no bead or antibody contact with target cells. The no-RBC-lysis protocol saves time and preserves cell viability. Isolated NK cells are untouched in their native, unactivated state — immediately ready for ADCC assays, tumor killing studies, cytokine profiling, innate immunity research, flow cytometry, scRNA-seq, and CAR-NK development.

Sorting Purity
Up to 95%
Sorting Time
18–20 min

Product Features of the Mouse NK Cell Sorting Kit

The Mouse NK Cell Sorting Kit delivers industry-leading performance for rapid CD49b+ NK 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-NK cells only, preserving NK cells in their native, unactivated state
Purity up to 95% — High-efficiency negative selection for highly pure NK cell populations
18–20 min fast sorting — Rapid protocol from mouse spleen to purified NK 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 NK Cell Sorting Kit uses column-free negative selection to isolate untouched CD49b+ NK 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 NK Cell Sorting Kit Works

Simple 4-step protocol for rapid cell isolation

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Step 1 of NK cells sorting: Sample Preparation - Grind mouse spleen, filter through 70 μm sterile nylon mesh, no RBC lysis, resuspend in FACS buffer at 1×10⁸ cells/mL for CS-0041

Step 1: Sample Preparation

Grind mouse spleen and filter through a 70 μm sterile nylon mesh to prepare a single-cell suspension (no RBC lysis needed). Count cells and resuspend in FACS buffer at 1×10⁸ cells/mL.

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Step 2 of NK cells sorting: Label Unwanted Cells - NK Cell Isolation Cocktail (100 μL/mL) at room temperature for 10 min, antibodies bind non-NK cells only with CS-0041

Step 2: Label Unwanted Cells

Add NK Cell Isolation Cocktail (100 μL/mL) to the sample and incubate at room temperature for 10 min. Biotin-conjugated antibodies bind exclusively to non-NK cells, leaving target NK cells completely untouched.

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Step 3 of NK cells sorting: Bind Labeled Cells to Magnetic Beads - Streptavidin Nanobeads (100 μL/mL), RT 5 min, top up with FACS buffer to 2.5 mL for CS-0041

Step 3: Bind Labeled Cells to Magnetic Beads

Vortex Streptavidin Nanobeads for 5–30 s, add to the sample (100 μL/mL), mix gently, and incubate at room temperature for 5 min. Then top up with FACS buffer to the working volume (e.g., 2.5 mL for a 5 mL tube).

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Step 4 of NK cells sorting: Magnetic Separation & Collection - Magnet 3 min single/5 min multi-position, tilt and pour off purified untouched NK cells for CS-0041

Step 4: Magnetic Separation & Collection

Place the tube on the magnet (3 min for a single-position magnet; 5 min for a multi-position magnet). Tilt the magnet and pour the supernatant into a new collection tube — the untouched NK cells are isolated and ready for downstream use.

Applications for the Mouse NK Cell Sorting Kit

This Mouse NK Cell Sorting Kit supports a wide range of downstream applications for CD49b+ NK 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 NK 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 NK Cell Sorting Kit contains everything needed for high-purity column-free negative selection from human PBMCs.

  • Mouse NK Cell Isolation Cocktail
  • Streptavidin Nanobeads
  • User manual

Storage & Shelf Life

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

Mouse NK Cell Sorting Kit — Frequently Asked Questions

Everything you need to know about the CS-0041 Mouse NK Cell Sorting Kit

Click each question to expand the answer

The CS-0041 Mouse NK Cell Sorting Kit from Cell Sorts uses column-free negative selection. NK Cell Isolation Cocktail (100 μL/mL) labels non-NK cells (T cells, B cells, monocytes, granulocytes, erythrocytes) at room temperature for 10 min. Streptavidin Nanobeads (100 μL/mL) bind labeled cells at room temperature for 5 min. A magnet (3 min single-position; 5 min multi-position) depletes non-targets; purified NK cells are poured off into a collection tube. No RBC lysis is required — no beads or antibodies ever contact the target cells.
Purity of isolated NK cells typically reaches up to 95%, as validated by flow cytometry using CD49b-APC (Clone DX5), CD3-FITC, NK1.1-PE, CD19-PerCP-Cy5.5 staining. The optimized negative selection protocol with NK Cell Isolation Cocktail and Streptavidin Nanobeads efficiently depletes all non-target cell populations while maximizing recovery of viable, functional NK cells from mouse spleen.
The CS-0041 kit is optimized for mouse spleen. Grind the spleen and filter through a 70 μm sterile nylon mesh to prepare a single-cell suspension (no RBC lysis needed). Count cells and resuspend in FACS buffer at 1×10⁸ cells/mL. The kit is also compatible with lymph node samples. Compatible with C57BL/6, BALB/c and other common mouse strains used in immunology research.
Step 1: Sample Preparation. Grind mouse spleen and filter through a 70 μm sterile nylon mesh (no RBC lysis needed). Count cells and resuspend in FACS buffer at 1×10⁸ cells/mL.
Step 2: Label Unwanted Cells. Add NK Cell Isolation Cocktail (100 μL/mL), incubate at room temperature for 10 min. Antibodies bind non-NK cells only.
Step 3: Bind Labeled Cells to Magnetic Beads. Vortex Streptavidin Nanobeads 5–30 s, add (100 μL/mL), mix gently, incubate room temperature for 5 min. Top up with FACS buffer to working volume (e.g., 2.5 mL for 5 mL tube).
Step 4: Magnetic Separation & Collection. Place on magnet (3 min single-position; 5 min multi-position). Tilt magnet and pour supernatant into new collection tube — untouched NK cells are isolated.
Total time: 18–20 min.
Isolated NK cells are immediately ready for: ADCC assays, tumor killing studies, cytokine profiling, innate immunity research, CAR-NK 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 NK 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 NK Cell Isolation Cocktail (100 μL/mL) targets CD3, CD4, CD8a, CD19, CD11b, Ly-6G, Ter-119 on non-target cells—effectively labeling T cells, B cells, monocytes, granulocytes, erythrocytes for depletion while leaving NK cells (CD49b+ NK1.1+) completely untouched. This comprehensive marker panel ensures up to 95% purity.
We recommend flow cytometry using CD49b-APC (Clone DX5), CD3-FITC, NK1.1-PE, CD19-PerCP-Cy5.5 antibodies. Typical results show up to 95% NK cells (CD49b+ CD3−) 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 NK 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-0041 uses column-free negative selection (18–20 min) with no RBC lysis required, achieving up to 95% purity with maximum speed and convenience. Column-based kits (25–35 min) offer higher consistency for larger sample sizes. Both yield untouched, bead-free NK cells. Choose CS-0041 for rapid processing with no RBC lysis; choose column-based for larger-scale isolation.

Related topics covered:

column-free negative selection CD49b+ NK Cell isolation CD49b+ NK Cell purity verification CD49b+ NK Cell sorting protocol column-free isolation single-cell analysis (scRNA-seq) CD49b+ NK Cell isolation markers PBMC CD49b+ NK Cell sorting

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