Human CD66b+ Neutrophil Isolation Kit CS-0009 | Cell Sorts™
CS-0009

Human Neutrophil Sorting Kit

Column-Free Positive Human

Cell Sorts™ Human Neutrophil Sorting Kit (CS-0009) enriches CD66b+ cells from erythrocyte-lysed human peripheral blood by column-free positive selection. A CD66b capture antibody labels target cells, prewashed releasable magnetic beads retain the labeled fraction on a compatible tube magnet, and release buffer detaches beads before final magnetic clarification. The recovered cells are bead-depleted, and the workflow does not require a separation column.

Supplied technical documentation specifies 15-minute antibody and bead incubations at 4°C, repeated 5-minute magnetic capture and wash steps, two 10-minute room-temperature release incubations, final magnetic clarification, and a 500 × g collection spin. In one representative flow-cytometry example, CD66b+ cells increased from 52.62% to 99.67%. Because CD66b is also expressed on eosinophils, laboratories should confirm population identity with an appropriate multicolor panel. Formats support up to 1×108, 5×108, or 1×109 starting cells. For research use only.

Sorting Purity
Representative result 99.67%
Sorting Time
Approximately 90 min after sample preparation

Product Features of the Human Neutrophil Sorting Kit

CS-0009 uses a documented column-free positive-selection workflow to enrich CD66b+ cells from erythrocyte-lysed human peripheral blood and remove magnetic beads before final collection.

Column-free positive selection - CD66b capture antibody and releasable magnetic beads enrich the labeled target fraction without a separation column
Bead-depleted recovery - Two release cycles and final magnetic clarification are specified before collecting the target-cell fraction
Defined starting material - The documented workflow begins with fresh anticoagulated human peripheral blood followed by red blood cell lysis
Representative 99.67% CD66b+ result - One supplied flow-cytometry example increased CD66b+ cells from 52.62% to 99.67%; results are sample- and workflow-dependent
Scalable formats - Kit configurations support up to 1×10⁸, 5×10⁸, or 1×10⁹ starting cells with proportional reagent volumes
Marker-aware interpretation - CD66b is also expressed on eosinophils, so post-sort identity should be confirmed with an appropriate multicolor panel

CS-0009 positively enriches CD66b+ cells from erythrocyte-lysed human peripheral blood. The supplied protocol includes repeated magnetic washes, two release cycles, final magnetic clarification, and a collection centrifugation; the listed timed steps total approximately 90 minutes after sample preparation.

How the Human Neutrophil Sorting Kit Works

Four practical stages summarizing the documented multi-step workflow

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Step 1 of CS-0009: prepare erythrocyte-lysed human peripheral-blood cells at 1×10⁸ cells per mL

Step 1: Prepare the Blood-Cell Suspension

Lyse red blood cells from fresh anticoagulated human blood, wash at 500 × g for 5 minutes, and resuspend at 1×10⁸ cells/mL in sterile-filtered PBS containing 2 mM EDTA and 2% FBS.

2
Step 2 of CS-0009: label CD66b-positive cells and bind prewashed releasable magnetic beads at 4°C

Step 2: Label CD66b+ Cells and Bind Beads

For 5×10⁷ cells in 500 µL, add 10 µL CD66b capture antibody and incubate at 4°C for 15 minutes. Add 100 µL prewashed releasable magnetic beads and incubate at 4°C for 15 minutes.

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Step 3 of CS-0009: capture and wash the labeled fraction on a compatible tube magnet

Step 3: Magnetically Capture and Wash

Bring the suspension to 2.5 mL, mix by pipetting, and place it on a compatible tube magnet for 5 minutes. Discard the supernatant and perform the documented magnetic wash sequence without allowing the beads to dry.

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Step 4 of CS-0009: perform two bead-release cycles, final magnetic clarification, and cell collection

Step 4: Release, Clarify, and Collect

Perform two 10-minute room-temperature release incubations, each followed by a 5-minute magnetic hold. Combine recovered supernatants, magnetically clarify for 5 minutes, then collect cells at 500 × g for 5 minutes.

Applications for the Human Neutrophil Sorting Kit

Potential downstream uses for the enriched CD66b+ fraction are listed below. Confirm post-sort identity, purity, and viability, and qualify each assay under laboratory-specific conditions.

Flow Cytometric Quality Control - Quantify CD66b+ enrichment, viability, and residual cell populations with a laboratory-validated multicolor 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 fraction in controlled studies after fit-for-purpose functional qualification Molecular Profiling - Prepare material for RNA, protein, or targeted molecular analysis while accounting for ex vivo handling effects Ex Vivo Stimulation Studies - Evaluate responses to defined stimuli after post-sort count, viability, and purity checks Sample-Preparation Method Development - Optimize red-cell lysis, magnetic washing, bead release, and downstream QC workflows Inflammation and Host-Response Research - Support hypothesis-driven studies using appropriately controlled CD66b+ cell preparations Drug-Response Research - Evaluate candidate compounds in laboratory-validated assays after confirming cell identity and quality

In addition, the Human Neutrophil Sorting Kit is compatible with a wide range of downstream applications, making it a versatile tool for your immunology research workflow.

Kit Components

CS-0009 includes the documented CD66b capture antibody, releasable magnetic beads, and release buffer for positive selection from erythrocyte-lysed human peripheral blood.

  • CD66b Capture Antibody
  • Releasable Magnetic Beads
  • Release Buffer

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-0009 Human Neutrophil Sorting Kit

Click each question to expand the answer

CS-0009 enriches a CD66b-positive cell fraction from erythrocyte-lysed human peripheral blood. CD66b is commonly used for human neutrophil enrichment, but it is also expressed on eosinophils; confirm the recovered population with a laboratory-validated multicolor panel.
The kit uses column-free positive selection. A CD66b capture antibody labels target cells, releasable magnetic beads retain the labeled fraction on a compatible tube magnet, and release buffer detaches the beads before final magnetic clarification.
The supplied documentation specifies fresh anticoagulated human whole blood, with EDTA anticoagulation recommended. Red blood cells are lysed, the remaining cells are washed at 500 × g for 5 minutes, and the suspension is adjusted to 1×108 cells/mL.
In one representative flow-cytometry example, the CD66b-positive population increased from 52.62% before separation to 99.67% after separation. This is an example result rather than a guaranteed specification; donor, sample handling, red-cell lysis, cell count, magnet performance, and gating can affect the outcome.
The manual lists 15-minute antibody and bead incubations, repeated 5-minute magnetic capture and wash steps, two 10-minute release incubations, additional 5-minute magnetic clarifications, and a 5-minute collection spin. Summing the listed timed steps gives approximately 90 minutes after sample preparation; red-cell lysis, counting, transfers, and QC add time.
The documented protocol performs two room-temperature release incubations and combines the recovered supernatants. A final magnetic hold removes residual magnetic material before the target-cell suspension is transferred for the 500 × g collection spin.
The manual specifies PBS containing 2 mM EDTA and 2% fetal bovine serum, sterile-filtered through a 0.22 µm membrane. Laboratories should prepare and qualify the buffer under their own procedures.
Documented formats support up to 1×108, 5×108, or 1×109 starting cells. The corresponding listed amounts are 20/100/200 µL capture antibody, 200 µL/1 mL/2×1 mL releasable beads, and 4/20/40 mL release buffer.
Use flow cytometry with a validated gating strategy and include viability and exclusion markers appropriate to the study. The supplied example used an Alexa Fluor 647 anti-CD66b reagent (clone 6/40c). Because CD66b alone does not distinguish every granulocyte subset, add markers suited to the sample and research question.
Store all kit components at 2-8°C and do not freeze. Refer to the component labels for expiry information, use a compatible magnetic rack, and follow the current product instructions. CS-0009 is for research use only and is not intended for diagnostic or therapeutic use.

Related topics covered:

column-free positive selection CD66b+ Cell (Neutrophil-Enriched Fraction) isolation CD66b+ Cell (Neutrophil-Enriched Fraction) purity verification CD66b+ Cell (Neutrophil-Enriched Fraction) sorting protocol column-free isolation post-sort flow-cytometric quality control CD66b+ Cell (Neutrophil-Enriched Fraction) isolation markers erythrocyte-lysed peripheral-blood cell sorting

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