CS-0002

Human B Cell Sorting Kit

Column-Based Negative Human

Cell Sorts™ Human B Cell Sorting Kit (CS-0002) enriches untouched CD19+ B cells from human PBMCs by column-based negative selection. Fc receptor blocking helps reduce nonspecific binding, while a biotin-antibody cocktail labels non-B cells and streptavidin ultra-nanobeads magnetically mark the non-target fraction. A magnetized separation column retains labeled cells, and unlabeled B cells are recovered in the gravity flow-through without direct bead labeling.

Supplied technical documentation specifies a 10-minute room-temperature antibody incubation, a 400×g wash, a 5-minute room-temperature nanobead incubation, and column equilibration and washing with 3 mL FACS buffer. One representative flow-cytometry example increased CD19+ cells from 9.78% before separation to 98.7% after sorting. Kit formats support up to 1×10⁸, 5×10⁸, or 1×10⁹ total cells. For research use only.

Sorting Purity

Representative result 98.7%

Sorting Time

Documented 10 + 5 min incubations

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Product Features of the Human B Cell Sorting Kit

CS-0002 uses a documented column-based negative-selection workflow to enrich untouched human CD19+ B cells from density-gradient-prepared PBMCs.

Column-based negative selection — Magnetically labeled non-B cells are retained while the target fraction is recovered by gravity flow

Untouched CD19+ target fraction — Recovered B cells are not directly labeled with antibodies or magnetic beads

Representative 98.7% post-sort purity — A documented example result, not a guaranteed value for every donor or sample

Defined room-temperature labeling — 10-minute antibody incubation followed by a documented wash and 5-minute nanobead incubation

Scalable kit formats — Options support up to 1×10⁸, 5×10⁸, or 1×10⁹ total starting cells

Controlled column recovery — 3 mL equilibration and 3 mL wash support consistent flow-through collection

CS-0002 uses column-based negative selection to enrich untouched human CD19+ B cells from PBMCs. The technical documentation specifies 10-minute antibody labeling, a 400×g wash for 5 minutes, 5-minute nanobead binding, and defined column-equilibration and wash steps, but it does not state a validated total processing time.

How the Human B Cell Sorting Kit Works

Simple 4-step protocol for rapid cell isolation

Step 1: Prepare and Count PBMCs

Prepare human PBMCs by density-gradient separation, count viable cells, and resuspend 1×10⁷ cells in 100 µL FACS buffer. Scale cell and reagent volumes proportionally.

Step 2: Block Fc Receptors and Label Non-B Cells

Add 10 µL Human FcR Blocker and 10 µL Human B Cell Isolation Cocktail per 1×10⁷ cells. Mix gently and incubate for 10 minutes at room temperature.

Step 3: Wash and Add Ultra-Nanobeads

Wash with ten times the cell-suspension volume of FACS buffer, centrifuge at 400×g for 5 minutes, restore to 100 µL, then add 10 µL well-mixed streptavidin ultra-nanobeads and incubate for 5 minutes at room temperature.

$Step 4 of CS-0002 protocol: use an equilibrated L column and collect untouched CD19+ B cells in the gravity flow-through and wash fraction$

Step 4: Separate and Collect Untouched B Cells

Equilibrate an L column with 3 mL FACS buffer. Bring the labeled sample to at least 500 µL, load it, collect the gravity flow-through, then wash with another 3 mL and combine the target-cell fractions.

Applications for the Human B Cell Sorting Kit

This Human B Cell Sorting Kit supports a wide range of downstream applications for CD19+ B Cell isolation and column-based negative selection workflows from human PBMCs.

Flow Cytometric Immunophenotyping — Assess post-sort CD19+ purity, viability, and residual non-B-cell populations with a laboratory-validated panel B Cell Activation and Proliferation — Study receptor-driven activation and proliferation after confirming post-sort cell quality Immunoglobulin Secretion Studies — Evaluate antibody secretion in appropriately controlled B-cell culture systems BCR Signaling Research — Investigate B-cell receptor signaling using an untouched target-cell fraction Cell Culture — Establish downstream culture workflows after post-sort count, viability, and sterility assessment Transcriptomic and BCR Repertoire Analysis — Prepare input for RNA, BCR-sequencing, or single-cell workflows when platform-specific QC criteria are met Autoimmune, Infectious-Disease, and Vaccine Research — Study human B-cell responses in laboratory-validated research models Drug Discovery — Evaluate candidate immunomodulators in appropriately controlled B-cell assays

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

Human FcR Blocker
Human B Cell Isolation Cocktail
Streptavidin Ultra-Nanobeads

Storage & Shelf Life

Storage

Store refrigerated at 4°C; do not freeze

Shelf Life

Refer to the product label

Human B Cell Sorting Kit — Frequently Asked Questions

Everything you need to know about the CS-0002 Human B Cell Sorting Kit

Click each question to expand the answer

CS-0002 is documented for enriching untouched CD19+ B cells from human PBMCs. Prepare PBMCs by density-gradient separation and count viable cells before labeling. Red-blood-cell lysis is not the recommended starting-sample preparation in the supplied protocol.

Fc receptor blocking helps reduce nonspecific binding. A biotin-conjugated isolation cocktail labels non-B cells, and streptavidin ultra-nanobeads magnetically mark that labeled fraction. The magnetic column retains labeled non-target cells while unlabeled CD19+ B cells are recovered in the gravity flow-through.

The supplied technical material reports purity above 90% and up to 99% under tested conditions. In one representative flow-cytometry example, CD19+ cells increased from 9.78% before separation to 98.7% after sorting. This is an example result, not a guaranteed value for every donor or run.

The documented protocol specifies a 10-minute room-temperature antibody incubation, a 400×g wash for 5 minutes, and a 5-minute room-temperature nanobead incubation. It does not state a validated total processing time; total duration varies with PBMC preparation, counting, wash, column flow, collection, and QC.

Resuspend 1×10⁷ cells in 100 µL FACS buffer, then add 10 µL Human FcR Blocker and 10 µL Human B Cell Isolation Cocktail. After the documented wash and restoration to 100 µL, add 10 µL well-mixed streptavidin ultra-nanobeads. Scale all volumes proportionally with total cell number.

The protocol specifies FACS buffer composed of 1× PBS, 2% FBS, and 0.02% EDTA. Labeling incubations are performed at room temperature with gentle mixing. Maintain stable osmolality, use low-endotoxin reagents, and follow sterile handling appropriate to the downstream assay.

Fc receptor blocking helps reduce nonspecific antibody binding. After the 10-minute cocktail incubation, the protocol requires a tenfold-volume wash followed by 400×g for 5 minutes before nanobead labeling. Omitting or altering this step may affect background labeling, purity, and recovery.

A documented L separation column supports a maximum load of 1×10⁸ total cells. Equilibrate the column with 3 mL FACS buffer, bring the labeled sample to at least 500 µL before loading, and avoid exceeding the column capacity. Split larger inputs across multiple columns.

Collect the initial gravity flow-through, then wash the column with an additional 3 mL FACS buffer and combine the target-cell fractions. Determine viable-cell yield and assess CD19+ purity with a laboratory-validated flow-cytometry panel that includes a viability marker and relevant non-B-cell exclusion markers.

Review PBMC viability, cell aggregation, cell concentration, proportional reagent scaling, nanobead resuspension, incubation conditions, the 400×g wash, column equilibration, sample volume, flow behavior, and collection technique. Do not load more than 1×10⁸ cells per L column, and verify both post-sort purity and viable-cell recovery.