Human Neutrophil Negative Selection Kit CS-0010 | Cell Sorts™
CS-0010

Human Neutrophil Sorting Kit

Column-Free Negative Human

Cell Sorts™ Human Neutrophil Sorting Kit (CS-0010) enriches untouched human neutrophils by column-free negative selection. The documented workflow starts with anticoagulated whole blood; density-gradient handling, red-cell lysis, and low-speed cleanup create a granulocyte-rich suspension. The supplied antibody cocktail and magnetic beads label non-neutrophil cells, while the target fraction remains unlabeled and is recovered by two controlled magnetic pour-offs.

For 1×107 prepared cells, the protocol specifies 10 µL antibody cocktail for 5 minutes, 8 µL magnetic beads for 5 minutes, and two 3-minute magnetic separations—an approximately 16-minute core selection after sample preparation. In one representative CD16-PE/CD66b-FITC plot, the double-positive quadrant increased from 75.0% to 98.4%; this is not a guaranteed specification. Verify identity, purity, viability, and function for each downstream assay. For research and in vitro cell-culture use only.

Sorting Purity
Representative result 98.4%
Sorting Time
Approximately 16 min core selection after sample preparation

Product Features of the Human Neutrophil Sorting Kit

CS-0010 uses a documented column-free negative-selection workflow to enrich untouched human neutrophils from a granulocyte-rich suspension prepared from anticoagulated whole blood.

Column-free negative selection - The antibody cocktail and magnetic beads label non-neutrophil cells for removal without a separation column
Untouched target fraction - Neutrophils remain unlabeled and are recovered by two controlled magnetic pour-off steps
Defined whole-blood preparation - The documented workflow uses density-gradient handling, red-cell lysis, and low-speed cleanup to prepare a granulocyte-rich suspension
Approximately 16-minute core selection - Listed selection steps are 5 minutes for antibody labeling, 5 minutes for bead binding, and two 3-minute magnetic holds after sample preparation
Representative 98.4% CD16+/CD66b+ result - One supplied flow-cytometry plot increased the double-positive quadrant from 75.0% to 98.4%; results are sample- and workflow-dependent
Scalable configurations - Documented formats provide 100 µL or 1 mL each of antibody cocktail and magnetic beads and support up to 1×10⁹ total cells

CS-0010 uses column-free negative selection to enrich an untouched human neutrophil fraction from a granulocyte-rich suspension prepared from anticoagulated whole blood. The listed core-selection steps total approximately 16 minutes; density-gradient processing, red-cell lysis, centrifugation, counting, transfers, and QC are additional.

How the Human Neutrophil Sorting Kit Works

Simple 4-step protocol for rapid cell isolation

1
Step 1 of CS-0010: prepare a granulocyte-rich suspension from human anticoagulated whole blood

Step 1: Prepare the Granulocyte-Rich Suspension

Process human anticoagulated whole blood by density-gradient separation, retain the granulocyte-containing red-cell layer, lyse red cells on ice for 15 minutes, complete the documented 500 × g and 120 × g centrifugation steps, and adjust to 5×10⁷ cells/mL.

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Step 2 of CS-0010: label non-neutrophil cells with 10 µL antibody cocktail for 5 minutes at 2-8°C

Step 2: Label Non-Neutrophil Cells

Transfer 1×10⁷ prepared cells to a 5 mL flow tube, add 10 µL antibody cocktail, mix, and incubate at 2-8°C for 5 minutes.

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Step 3 of CS-0010: add 8 µL magnetic beads and incubate for 5 minutes at 2-8°C

Step 3: Bind Magnetic Beads

Add 8 µL thoroughly mixed magnetic beads, mix gently, incubate at 2-8°C for 5 minutes, then add 2 mL sorting buffer and mix by pipetting.

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Step 4 of CS-0010: perform two 3-minute magnetic holds and controlled pour-offs

Step 4: Perform Two Magnetic Pour-Offs

Place the tube on a compatible 5 mL tube magnet for 3 minutes and pour the enriched suspension into a fresh tube. Repeat the 3-minute magnetic hold and controlled pour-off once more to collect the untouched target fraction.

Applications for the Human Neutrophil Sorting Kit

Potential downstream uses for the enriched untouched neutrophil fraction are listed below. Confirm identity, purity, viability, and functional suitability under laboratory-specific conditions.

Flow Cytometric Quality Control - Measure CD16/CD66b enrichment, viability, and residual populations with a laboratory-validated granulocyte panel Neutrophil Phenotyping - Characterize surface-marker patterns after confirming that the recovered fraction meets assay-specific acceptance criteria Innate Immune Cell Biology - Use the enriched untouched fraction in controlled studies after fit-for-purpose functional qualification Molecular Profiling - Prepare material for DNA, RNA, or protein analysis while accounting for whole-blood processing and ex vivo handling Ex Vivo Stimulation Studies - Evaluate responses to defined stimuli after post-sort identity, count, viability, and purity checks Sample-Preparation Method Development - Optimize density-gradient handling, red-cell lysis, low-speed cleanup, magnetic separation, and downstream QC Inflammation and Host-Response Research - Support hypothesis-driven studies using appropriately controlled neutrophil-enriched preparations Drug-Response Research - Evaluate candidate compounds in laboratory-validated assays after confirming cell identity and quality

The listed applications are potential research uses rather than guaranteed performance claims. Confirm post-sort identity, purity, viability, and functional suitability under laboratory-specific conditions.

Kit Components

CS-0010 includes the documented human neutrophil negative-selection antibody cocktail and magnetic beads for use with a prepared whole-blood granulocyte fraction.

  • Human Neutrophil Negative Selection Antibody Cocktail
  • Human Neutrophil Negative Selection Magnetic Beads

Storage & Shelf Life

Storage
Store at 2-8°C; do not freeze
Shelf Life
Refer to the product label

Human Neutrophil Sorting Kit — Frequently Asked Questions

Everything you need to know about the CS-0010 Human Neutrophil Sorting Kit

Click each question to expand the answer

CS-0010 enriches an untouched human neutrophil fraction from a granulocyte-rich suspension prepared from anticoagulated whole blood. The target fraction is not directly labeled by the antibody cocktail or magnetic beads.
The kit uses column-free negative selection. Non-neutrophil cells are labeled with the supplied antibody cocktail and magnetic beads, retained on a compatible tube magnet, and separated from the poured-off target fraction.
The supplied documentation starts with human anticoagulated whole blood. It describes density-gradient separation, retention of the red-cell layer containing granulocytes, red-cell lysis on ice, centrifugation, and adjustment of the prepared suspension to 5×107 cells/mL.
After the granulocyte-rich suspension has been prepared, the listed steps are a 5-minute antibody incubation, a 5-minute bead incubation, and two 3-minute magnetic holds. These timed steps total approximately 16 minutes. Density-gradient processing, red-cell lysis, centrifugation, counting, transfers, and QC require additional time.
For 1×107 prepared cells, the manual specifies 10 µL antibody cocktail followed by 8 µL magnetic beads. Reagent volumes should be scaled proportionally above 1×107 cells; samples below that number still use the listed single-reaction amounts.
In one representative CD16-PE/CD66b-FITC flow-cytometry plot, the double-positive quadrant increased from 75.0% before separation to 98.4% after separation. This is an example result rather than a guaranteed product specification; sample handling, lysis, counting, mixing, magnet performance, and gating can affect the outcome.
The supplied manual describes the cocktail as targeting non-neutrophil surface markers, but it does not enumerate the individual antibody specificities. Use the current lot documentation or contact technical support when exact cocktail composition is required for experimental interpretation.
Use a laboratory-validated granulocyte gating strategy with viability and exclusion controls appropriate to the study. The supplied example used CD16-PE and CD66b-FITC. Confirm purity, identity, and recovery under the laboratory’s own sample-preparation and instrument settings.
The sorting buffer is PBS at pH 7.2 containing 0.5% BSA and 2 mM EDTA; the manual notes that HSA, human serum, or FBS may replace BSA and sodium citrate may replace EDTA. Store kit components at 2-8°C, do not freeze, and refer to the product label for expiry.
The supplied documentation lists flow cytometry, culture, and DNA/RNA extraction as potential uses. For stimulation, host-response, molecular, or drug-response studies, first confirm post-sort identity, purity, viability, and functional suitability. CS-0010 is for research and in vitro cell-culture use only, not for diagnostic or therapeutic use.

Related topics covered:

column-free negative selection Untouched Human Neutrophils (CD16+/CD66b+ Enriched Fraction) isolation Untouched Human Neutrophils (CD16+/CD66b+ Enriched Fraction) purity verification Untouched Human Neutrophils (CD16+/CD66b+ Enriched Fraction) sorting protocol column-free isolation post-sort CD16/CD66b flow-cytometric quality control Untouched Human Neutrophils (CD16+/CD66b+ Enriched Fraction) isolation markers whole-blood granulocyte preparation and sorting

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