CS-0035

Mouse B Cell Sorting Kit

Column-Free Negative Mouse

Sorting Purity

Up to 99%

Sorting Time

18–20 min

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Product Features of the Mouse B Cell Sorting Kit

The Mouse B Cell Sorting Kit delivers industry-leading performance for rapid CD19+ B Cell isolation from PBMCs via column-free negative selection.

Column-free separation — No magnetic column needed; streamlined workflow isolates untouched B cells directly from mouse splenocytes and lymph node cell suspensions by simple pour-off collection

Zero contact with target cells — Antibodies and streptavidin nanobeads bind only non-B cells; B cells remain completely untouched in their native, unactivated state

Purity 90-99% — High-efficiency negative selection delivers ultra-pure B cells populations, validated by flow cytometry

18-20 min fast protocol — Rapid column-free isolation from sample to purified B cells, saving valuable research time

No bead residue — Isolated B cells are completely bead-free and ready for immediate downstream applications including flow cytometry, cell culture, functional assays, CRISPR gene editing, scRNA-seq, BCR repertoire sequencing

Easy handling — Minimal steps, user-friendly procedure suitable for any lab; no expensive FACS equipment or specialized training required

The Mouse B Cell Sorting Kit uses column-free negative selection to isolate untouched CD19+ B 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 B Cell Sorting Kit Works

Simple 4-step protocol for rapid cell isolation

Step 1: Sample Preparation

Prepare single-cell suspension from mouse splenocytes and lymph node cell suspensions. Pass through 70 μm strainer, ACK lysis if needed. Count cells, resuspend at 1×10⁸ cells/mL in separation buffer (2–8°C).

Step 2: Antibody Cocktail Labeling

Add 10 μL Mouse B Cell Isolation Cocktail per 1×10⁷ cells. Mix, incubate 4°C 5–10 min. Labels non-B cells (T cells, NK cells, monocytes, granulocytes); B cells untouched.

Step 3: Magnetic Nanobead Binding

Vortex Streptavidin Nanobeads, add 10 μL per 1×10⁷ cells. Mix, incubate 4°C 5–10 min. Nanobeads bind labeled non-target cells.

Step 4: Column-Free Separation & Collection

Dilute to 2.5 mL, place on magnetic separator 3–5 min. Pour off supernatant containing purified B cells — ready for B cell activation, antibody production, BCR signaling, hybridoma generation, vaccine development.

Applications for the Mouse B Cell Sorting Kit

This Mouse B Cell Sorting Kit supports a wide range of downstream applications for CD19+ B Cell isolation and column-free negative selection workflows from mouse splenocytes or lymph node cells.

B Cell Biology Research — B cell activation, differentiation, proliferation, and antibody production studies in mouse models Humoral Immunity Studies — Antigen-specific B cell response and immunoglobulin repertoire analysis Autoimmune Disease Models — B cell-mediated autoimmunity mechanism research (e.g., SLE, lupus mouse models) Infectious Disease Research — Pathogen-specific B cell response and neutralizing antibody development Flow Cytometry — High-purity B cell populations ready for CD19/CD45R(B220) staining and analysis Cell Culture — Unactivated, bead-free B cells ideal for in vitro expansion and manipulation Drug Discovery — B cell-targeted drug screening in syngeneic and transgenic mouse models Adoptive Transfer — Highly viable, naive-state B cells for mouse adoptive cell transfer experiments

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

Mouse CD16/CD32 Blocker
Mouse B Cell Isolation Cocktail
Streptavidin Nanobeads
User manual

Storage & Shelf Life

Storage

Store at 2-8°C protected from light

Shelf Life

12 months

Mouse B Cell Sorting Kit — Frequently Asked Questions

Everything you need to know about the CS-0035 Mouse B Cell Sorting Kit

Click each question to expand the answer

The CS-0035 Mouse B Cell Sorting Kit from Cell Sorts uses column-free negative selection. Non-target cells (T cells, NK cells, monocytes, granulocytes) are labeled with a biotin-conjugated antibody cocktail and removed by magnetic separation, while untouched B cells remain in the supernatant. This ensures no beads or antibodies ever contact the target cells, preserving their native functional state.

Purity of isolated B cells typically reaches 90-99%, as validated by flow cytometry using CD19-PE (Clone 6D5), B220-APC, CD3-FITC staining. The optimized antibody cocktail efficiently depletes all non-target cell populations while maximizing recovery of viable, functional B cells from mouse splenocytes and lymph node cell suspensions.

The CS-0035 kit is optimized for mouse splenocytes and lymph node cell suspensions. Prepare a single-cell suspension by gentle mechanical disruption through a 70 μm cell strainer. Perform red blood cell lysis if needed (e.g., ACK lysis buffer). The kit is compatible with C57BL/6, BALB/c and other common mouse strains used in immunology research.

Step 1: Prepare a single-cell suspension from mouse splenocytes and lymph node cell suspensions.
Step 2: Count cells and resuspend at recommended density.
Step 3: Incubate with Mouse B Cell Isolation Cocktail at 4°C for 5–10 min.
Step 4: Add Streptavidin Nanobeads and incubate at 4°C for 5–10 min.
Step 5: Place on magnetic separator for 3–5 min.
Step 6: Pour off purified B cells into collection tube.
Total time: 18-20 min.

Isolated B cells are immediately ready for: B cell activation, antibody production, BCR signaling, hybridoma generation, 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 B 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 CD3, CD4, CD8a, CD11b, CD49b, Ly-6G on non-target cells—effectively labeling T cells, NK cells, monocytes, granulocytes for depletion while leaving B cells completely untouched. This comprehensive marker panel ensures high purity and recovery of the target cell population.

We recommend flow cytometry using CD19-PE (Clone 6D5), B220-APC, CD3-FITC antibodies. Typical results show 90-99% target cells 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 B 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-0035 uses column-free negative selection (18-20 min), ideal for speed and convenience with smaller samples. Column-based kits (25–35 min) offer higher consistency for larger sample sizes. Both yield untouched, bead-free B cells with 90-99% purity. Choose CS-0035 for rapid processing; choose column-based for larger-scale isolation.