CS-0042

Mouse NK Cell Sorting Kit

Column-Based Negative Mouse

The Cell Sorts CS-0042 Mouse NK Cell Sorting Kit uses column-assisted negative selection to isolate untouched NK cells from mouse spleen with 65-92% purity and high recovery. Mouse NK Cell Isolation Cocktail (10 μL) labels non-NK cells at room temperature for 10 min. After wash/centrifugation (500×g, 5 min), Ultra-Nanobeads (10 μL) bind at room temperature for 5 min. An L separation column depletes non-targets by gravity drip-through; 3 mL FACS Buffer wash maximizes yield.

No bead or antibody contact with target cells. The column-based format handles larger cell numbers with consistent performance. Isolated NK cells are bead-free and 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

65–92%

Sorting Time

25–35 min

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Product Features of the Mouse NK Cell Sorting Kit

The Mouse NK Cell Sorting Kit delivers industry-leading performance for rapid NK Cell (CD49b+) 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 NK cells are bead-free and ready for immediate downstream use

Purity 65-92% — Reliable negative selection for enriched NK cell populations

High recovery rate — Optimized column format maximizes yield of viable NK cells

Suitable for large samples — Column-based format handles larger cell numbers with consistent performance

Native state preservation — Antibodies and beads do not contact NK cells, maintaining their original cytotoxic function

The Mouse NK Cell Sorting Kit uses column-based negative selection to isolate untouched NK Cells (CD49b+) 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 NK Cell Sorting Kit Works

Simple 4-step protocol for rapid cell isolation

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).

Step 2: Antibody Labeling

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

Step 3: Magnetic Bead Binding & Wash

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

Step 4: Column-Based Separation & Collection

Place an L separation column in the magnetic field, equilibrate with 3 mL FACS Buffer. Load cell suspension, collect NK cells as they drip through by gravity. Add another 3 mL FACS Buffer to wash and collect residual cells for maximum yield.

Applications for the Mouse NK Cell Sorting Kit

This Mouse NK Cell Sorting Kit supports a wide range of downstream applications for NK Cell (CD49b+) isolation and column-based 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 — Enriched NK cell populations ready for CD49b/NK1.1 staining and analysis Cell Culture — Bead-free, unactivated 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-based negative selection from human PBMCs.

Mouse NK Cell Isolation Cocktail
Ultra-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-0042 Mouse NK Cell Sorting Kit

Click each question to expand the answer

The CS-0042 Mouse NK Cell Sorting Kit from Cell Sorts uses column-assisted negative selection. Mouse NK Cell Isolation Cocktail (10 μL) labels non-NK cells (T cells, B cells, monocytes, granulocytes, erythrocytes) at room temperature for 10 min. After wash/centrifugation (500×g, 5 min), Ultra-Nanobeads (10 μL) bind labeled cells at room temperature for 5 min. An L separation column in a magnetic field depletes non-targets by gravity drip-through. NK cells pass through untouched — bead-free and ready for downstream use.

Purity of isolated NK cells typically reaches 65–92%, as validated by flow cytometry using CD49b-APC (Clone DX5), CD3-FITC, NK1.1-PE, CD19-PerCP-Cy5.5 staining. The column-assisted format with wash/centrifugation step efficiently depletes non-NK cells while maximizing recovery of viable NK cells. Note: NK cell proportions vary by mouse strain and tissue; purity within the 65–92% range is normal for enriched NK cell populations.

The CS-0042 kit is optimized for mouse spleen. Grind the 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). The kit is also compatible with lymph node samples. Compatible with C57BL/6, BALB/c and other common mouse strains.

Step 1: Sample Preparation. Grind mouse spleen through a 70 μm sterile nylon mesh filter. Count cells and resuspend 1×10⁷ cells in 100 μL FACS Buffer.
Step 2: Antibody Labeling. Add 10 μL Mouse NK Cell Isolation Cocktail, mix gently by pipetting 2–3 times, incubate at room temperature for 10 min to label non-NK cells.
Step 3: Magnetic Bead Binding & Wash. Wash cells with 10× volume of FACS Buffer, centrifuge at 500×g for 5 min, resuspend in 100 μL. Vortex Ultra-Nanobeads 5–30 s, add 10 μL, mix gently, incubate RT for 5 min.
Step 4: Column-Based Separation & Collection. Place L separation column in magnetic field, equilibrate with 3 mL FACS Buffer. Load cell suspension, collect NK cells as they drip through by gravity. Wash with 3 mL FACS Buffer for maximum yield.
Total hands-on time: ~25–30 min; total protocol: ~40–50 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. Bead-free and antibody-free output preserves full cytotoxic function.

Antibodies and Ultra-Nanobeads bind exclusively to non-target cells — they never touch NK cells. The untouched approach ensures full preservation of surface receptors (CD49b, NK1.1, NKG2D), native cytotoxic function, and physiological relevance for downstream assays. The L separation column provides gentle, efficient depletion without mechanical stress on target cells.

The Mouse NK Cell Isolation Cocktail (10 μL) 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 ensures 65–92% enriched NK cell purity.

We recommend flow cytometry using CD49b-APC (Clone DX5), CD3-FITC, NK1.1-PE, CD19-PerCP-Cy5.5 antibodies. Typical results show 65–92% 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-0042 uses column-assisted negative selection (~25–30 min hands-on) with an L separation column and Ultra-Nanobeads, achieving 65–92% purity with high recovery for larger samples. It includes a centrifugation step (500×g, 5 min) and wash for enhanced purity. The CS-0041 uses column-free negative selection (18–20 min) with Streptavidin Nanobeads, achieving up to 95% purity with maximum speed and no RBC lysis. Both preserve NK cell native cytotoxic function. Choose CS-0042 for larger-scale isolation with high recovery; choose CS-0041 for rapid processing.