CS-0033

Mouse Neutrophil Sorting Kit

Column-Free Negative Mouse

The Cell Sorts CS-0033 Mouse Neutrophil Sorting Kit uses column-free negative selection with FcR blocking to isolate untouched neutrophils from mouse bone marrow with >90% purity, up to 99% in 18–20 min. Mouse CD16/CD32 Blocker and Mouse Neutrophil Isolation Cocktail (100 μL/mL each) block Fc receptors and label non-neutrophil cells at 4°C for 15 min. After wash/centrifugation (400×g, 5 min), Streptavidin Nanobeads (100 μL/mL) bind at 4°C for 15 min. Magnetic separator 3–5 min, then pour off purified neutrophils.

No columns, no bead or antibody contact with target cells. FcR blocking prevents non-specific binding for ultra-high purity. Isolated neutrophils are untouched in their native, unactivated state — immediately ready for NETosis assays, phagocytosis studies, bacterial killing assays, inflammation research, flow cytometry, scRNA-seq, and innate immunity studies.

Sorting Purity

90-99%

Sorting Time

18–20 min

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Product Features of the Mouse Neutrophil Sorting Kit

The Mouse Neutrophil Sorting Kit delivers industry-leading performance for rapid Ly-6G+ Neutrophils isolation from PBMCs via column-free negative selection.

Column-free separation — No column needed, streamlined workflow; simply pour target cells into a new tube

Zero contact with target cells — Antibodies and beads bind non-mononuclear cells only, preserving neutrophils in their native, unactivated state

Purity >90%, up to 99% — High-efficiency negative selection for ultra-pure neutrophil populations

18–20 min fast sorting — Rapid protocol from mouse bone marrow to purified neutrophils

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 Neutrophil Sorting Kit uses column-free negative selection to isolate untouched Ly-6G+ Neutrophils 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 Neutrophil Sorting Kit Works

Simple 4-step protocol for rapid cell isolation

Step 1: Sample Preparation

Harvest mouse bone marrow from leg bones and prepare a single-cell suspension. Count cells and resuspend in pre-cooled FACS Buffer at 1×10⁸ cells/mL (e.g., 1×10⁷ cells in 100 μL).

Step 2: FcR Blocking & Antibody Labeling

Add Mouse CD16/CD32 Blocker (100 μL/mL) and Mouse Neutrophil Isolation Cocktail (100 μL/mL), mix gently by pipetting 2–3 times, and incubate at 4°C for 15 min to block Fc receptors and label non-neutrophil cells.

Step 3: Magnetic Bead Binding & Wash

Wash cells with 10× volume of pre-cooled FACS Buffer, centrifuge at 400×g for 5 min, and resuspend in the original volume. Vortex Streptavidin Nanobeads for 5–30 s, add 100 μL per mL of cell suspension, mix gently, and incubate at 4°C for 15 min.

Step 4: Column-Free Separation & Collection

Dilute with pre-cooled 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 neutrophils into a fresh collection tube.

Applications for the Mouse Neutrophil Sorting Kit

This Mouse Neutrophil Sorting Kit supports a wide range of downstream applications for Ly-6G+ Neutrophils isolation and column-free negative selection workflows from mouse splenocytes or lymph node cells.

Neutrophil Biology Research — Phagocytosis, chemotaxis, NETosis, and degranulation studies in mouse models Innate Immunity Studies — Host defense mechanisms and pathogen-neutrophil interactions Inflammation Research — Acute and chronic inflammation models, cytokine release assays Infectious Disease Research — Bacterial, fungal, and viral pathogen responses Autoimmune Disease Studies — Neutrophil extracellular trap (NET) formation in autoimmunity Cancer Immunology — Tumor microenvironment and tumor-associated neutrophil studies Drug Discovery — Neutrophil-targeted drug screening and functional assays

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

Mouse CD16/CD32 Blocker
Mouse Neutrophil Isolation Cocktail
Streptavidin Nanobeads
User manual

Storage & Shelf Life

Storage

Store at 2-8°C protected from light

Shelf Life

12 months

Mouse Neutrophil Sorting Kit — Frequently Asked Questions

Everything you need to know about the CS-0033 Mouse Neutrophil Sorting Kit

Click each question to expand the answer

The CS-0033 Mouse Neutrophil Sorting Kit from Cell Sorts uses column-free negative selection. Mouse CD16/CD32 Blocker (100 μL/mL) blocks Fc receptors, then Mouse Neutrophil Isolation Cocktail (100 μL/mL) labels non-neutrophil cells (T cells, B cells, NK cells, monocytes/macrophages, erythrocytes) at 4°C for 15 min. After wash/centrifugation (400×g, 5 min), Streptavidin Nanobeads (100 μL per mL) bind labeled cells at 4°C for 15 min. A magnetic separator (3–5 min) depletes non-targets; purified neutrophils are poured off into a collection tube. No beads or antibodies ever contact the target cells.

Purity of isolated neutrophils typically reaches >90%, up to 99%, as validated by flow cytometry using Ly-6G-APC (Clone 1A8), CD11b-FITC, F4/80-PE staining. The optimized negative selection protocol with FcR blocking, Mouse Neutrophil Isolation Cocktail, and Streptavidin Nanobeads efficiently depletes all non-target cell populations while maximizing recovery of viable, functional neutrophils from mouse bone marrow.

The CS-0033 kit is optimized for mouse bone marrow from leg bones. Harvest bone marrow, prepare a single-cell suspension, and resuspend in pre-cooled FACS Buffer at 1×10⁸ cells/mL (e.g., 1×10⁷ cells in 100 μL). The kit is also compatible with peripheral blood and peritoneal lavage. Compatible with C57BL/6, BALB/c and other common mouse strains used in immunology research.

Step 1: Sample Preparation. Harvest mouse bone marrow from leg bones and prepare a single-cell suspension. Count cells and resuspend in pre-cooled FACS Buffer at 1×10⁸ cells/mL (e.g., 1×10⁷ cells in 100 μL).
Step 2: FcR Blocking & Antibody Labeling. Add Mouse CD16/CD32 Blocker (100 μL/mL) and Mouse Neutrophil Isolation Cocktail (100 μL/mL), mix gently by pipetting 2–3 times, and incubate at 4°C for 15 min.
Step 3: Magnetic Bead Binding & Wash. Wash cells with 10× volume of pre-cooled FACS Buffer, centrifuge at 400×g for 5 min, and resuspend in the original volume. Vortex Streptavidin Nanobeads for 5–30 s, add 100 μL per mL of cell suspension, mix gently, and incubate at 4°C for 15 min.
Step 4: Column-Free Separation & Collection. Dilute with pre-cooled FACS Buffer to 2.5 mL (5 mL tube) or 7.5 mL (15 mL tube), place on the magnetic separator for 3–5 min, then pour off the untouched, purified neutrophils into a fresh collection tube.
Total time: 18–20 min.

Isolated neutrophils are immediately ready for: NETosis assays, phagocytosis studies, bacterial killing assays, inflammation research, innate immunity, 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 neutrophils. 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 Mouse Neutrophil Isolation Cocktail (100 μL/mL) targets CD3, CD4, CD8a, CD19, CD49b, Ter-119, F4/80 on non-target cells—effectively labeling T cells, B cells, NK cells, monocytes/macrophages, erythrocytes for depletion while leaving neutrophils (Ly-6G+ CD11b+) completely untouched. This comprehensive marker panel ensures >90% purity, up to 99%.

We recommend flow cytometry using Ly-6G-APC (Clone 1A8), CD11b-FITC, F4/80-PE antibodies. Typical results show >90%, up to 99% neutrophils (Ly-6G+ CD11b+) in the collected fraction. Viability can be assessed using Trypan blue exclusion or 7-AAD staining. For best results, gate on neutrophils by forward and side scatter, then analyze marker expression within the gated population.

Absolutely. Since isolated neutrophils 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-0033 uses column-free negative selection (18–20 min) with FcR blocking (CD16/CD32 Blocker), achieving >90% purity, up to 99% with maximum speed and convenience. Column-based kits (25–35 min) offer higher consistency for larger sample sizes. Both yield untouched, bead-free neutrophils. Choose CS-0033 for rapid processing with FcR-blocked purity; choose column-based for larger-scale isolation.