Human NK Cell Negative Selection Kit CS-0011 | Cell Sorts™
CS-0011

Human NK Cell Sorting Kit

Column-Free Negative Human

Cell Sorts™ Human NK Cell Sorting Kit (CS-0011) enriches untouched human NK cells from PBMCs by column-free negative selection. A biotin-conjugated antibody cocktail labels non-NK cells; streptavidin nanobeads bind the labeled fraction for retention on a tube magnet, while the unlabeled NK-cell fraction is collected by pour-off. The documented workflow uses pre-cooled FACS buffer and 4°C incubations to support controlled processing.

For 1×107 PBMCs, the protocol specifies 10 µL cocktail, a 15-minute incubation, a 400 × g wash, a lot-specific nanobead dose, a second 15-minute incubation, and 3–5 minutes of magnetic capture. One representative healthy-donor example increased CD3-/CD56+ NK cells from approximately 13% before separation to 94.5% ± 5% after separation; this is not a guaranteed specification. Confirm identity, purity, viability, recovery, and function for each sample and assay. For research use only.

Sorting Purity
Typical 85-92%; up to 95%
Sorting Time
Approximately 38-40 min timed steps after PBMC preparation

Product Features of the Human NK Cell Sorting Kit

CS-0011 uses a documented column-free negative-selection workflow to enrich untouched human NK cells from PBMCs using pre-cooled buffer, two 15-minute incubations, a 400 × g wash, and 3-5 minutes of magnetic capture.

Column-free negative selection - The antibody cocktail and streptavidin nanobeads label non-NK cells for magnetic retention without a separation column
Untouched target fraction - The unlabeled NK-cell fraction is recovered by controlled pour-off after magnetic capture of labeled cells
Defined PBMC workflow - The documentation specifies pre-cooled FACS buffer, 4°C incubations, a 400 × g wash, and magnetic separation
Approximately 38-40 minutes of timed steps - Two 15-minute incubations, a 5-minute wash, and a 3-5 minute magnetic hold are specified after PBMC preparation
Representative CD3-/CD56+ result - One healthy-donor example increased from approximately 13% before separation to 94.5% ± 5% after separation; results are sample- and workflow-dependent
Scalable formats with lot-specific dosing - Configurations support up to 1×10⁸, 5×10⁸, or 1×10⁹ cells; follow the current lot insert for the nanobead volume

The Human NK Cell Sorting Kit uses column-free negative selection to isolate untouched Untouched Human NK Cells (CD3-/CD56+ Enriched Fraction) from human PBMCs. Review the product-specific four-stage workflow below. The stated sorting time of Approximately 38-40 min timed steps after PBMC preparation covers the specified timed stages only; sample preparation, handling, optional collection, and quality control require additional time. Follow the current product instructions and validate each preparation before downstream use.

How the Human NK Cell Sorting Kit Works

Four stages summarizing the documented PBMC negative-selection workflow

1
Step 1 of CS-0011: prepare density-gradient human PBMCs in pre-cooled FACS buffer at no more than 1×10⁸ cells per mL

Step 1: Prepare PBMCs

Prepare human PBMCs by density-gradient separation, count cells, and resuspend in pre-cooled FACS buffer at no more than 1×10⁸ cells/mL. Do not use red-blood-cell lysis for PBMC preparation.

2
Step 2 of CS-0011: add 10 µL NK-cell isolation cocktail per 1×10⁷ cells and incubate for 15 minutes at 4°C

Step 2: Label Non-NK Cells

For 1×10⁷ cells in 100 µL, add 10 µL Human NK Cell Isolation Cocktail, mix gently, and incubate for 15 minutes at 4°C.

3
Step 3 of CS-0011: wash at 400 × g for 5 minutes, add the current lot-specific nanobead volume, and incubate for 15 minutes at 4°C

Step 3: Wash and Bind Nanobeads

Add 10 volumes of pre-cooled buffer, centrifuge at 400 × g for 5 minutes, and resuspend. Mix the streptavidin nanobeads for 5-30 seconds, use the current lot-specific dose, and incubate for 15 minutes at 4°C.

4
Step 4 of CS-0011: magnetically capture for 3-5 minutes and pour the untouched NK-cell fraction into a new tube

Step 4: Capture and Collect

Add at least 5 volumes of pre-cooled buffer, magnetically capture for 3 minutes in a single-tube separator or 5 minutes in a multi-tube rack, then pour the untouched NK-cell fraction into a new tube. Optional final collection is 400 × g for 10 minutes.

Applications for the Human NK Cell Sorting Kit

Potential downstream uses for the enriched untouched NK-cell fraction are listed below. Confirm CD3-/CD56+ identity, purity, viability, recovery, and functional suitability under laboratory-specific conditions.

Flow Cytometric Quality Control - Measure CD3-/CD56+ enrichment, viability, recovery, and residual populations with a laboratory-validated panel NK Cell Phenotyping - Characterize NK-cell surface markers only after confirming that the recovered fraction meets assay-specific acceptance criteria Controlled Cell Culture and Expansion - Establish culture conditions after post-sort identity, count, viability, and sterility assessment Cytotoxicity and Target-Cell Response Studies - Use qualified NK-cell preparations in controlled target-cell assays with appropriate controls Cytokine and Innate-Immune Response Studies - Evaluate responses to defined stimuli after confirming sample quality and functional suitability Molecular and Single-Cell Profiling - Prepare DNA, RNA, protein, or single-cell inputs when platform-specific quality requirements are met Sample-Preparation and Separation Method Development - Optimize PBMC preparation, buffer handling, reagent dosing, magnetic capture, and downstream QC Drug-Response Research - Evaluate candidate compounds in laboratory-validated assays after confirming cell identity and quality

The listed applications are potential research uses, not guaranteed performance claims. Confirm post-sort identity, purity, viability, recovery, and functional suitability under laboratory-specific conditions.

Kit Components

CS-0011 includes the documented Human NK Cell Isolation Cocktail and Streptavidin Nanobeads for column-free negative selection from human PBMCs.

  • Human NK Cell Isolation Cocktail
  • Streptavidin Nanobeads

Storage & Shelf Life

Storage
Store at 4°C; do not freeze
Shelf Life
Refer to the product label

Human NK Cell Sorting Kit — Frequently Asked Questions

Everything you need to know about the CS-0011 Human NK Cell Sorting Kit

Click each question to expand the answer

CS-0011 enriches an untouched human NK-cell fraction from human PBMCs. The representative flow-cytometry example identifies the enriched population as CD3-/CD56+.
The kit uses column-free negative selection. A biotin-conjugated cocktail labels non-NK cells, streptavidin nanobeads bind the labeled fraction, and a compatible tube magnet retains those cells while the unlabeled NK-cell fraction is poured into a new tube.
Use human PBMCs prepared by density-gradient separation, count the cells, and resuspend them in pre-cooled FACS buffer at no more than 1×108 cells/mL. The documentation advises against red-blood-cell lysis for PBMC preparation.
After PBMC preparation, the listed stages are a 15-minute cocktail incubation, a 5-minute 400 × g wash, a 15-minute nanobead incubation, and a 3-5 minute magnetic hold. These specified timed stages total approximately 38-40 minutes; counting, transfers, buffer preparation, optional collection, and QC require additional time.
For 1×107 cells in 100 µL, the dosing table specifies 10 µL Human NK Cell Isolation Cocktail followed by a 15-minute incubation at 4°C. Scale according to the current product instructions.
The supplied document contains a dosing inconsistency: the procedure lists 75 µL/mL, while the dosing table lists 10 µL per 1×107 cells in 100 µL. Use the current lot-specific Cell Sorts insert or contact technical support before running the assay rather than inferring a volume.
The documentation describes typical purity of approximately 85-92%, with results up to 95%. In one representative healthy-donor example, CD3-/CD56+ cells increased from approximately 13% to 94.5% ± 5%. These values are examples, not guaranteed specifications.
The supplied document describes an antibody cocktail that labels non-NK cells but does not enumerate the individual antibody specificities. Use the current lot documentation or contact technical support if exact cocktail composition is required for study interpretation.
Use pre-cooled FACS buffer consisting of 1× PBS, 2% FBS, and 0.02% EDTA. Store the cocktail and nanobeads at 4°C and do not freeze. After separation, verify CD3-/CD56+ identity together with purity, viability, and recovery using a laboratory-validated panel.
The documentation lists flow cytometry, cell culture, and functional assays as potential research uses. Molecular, single-cell, cytotoxicity, cytokine, and drug-response studies require fit-for-purpose qualification. CS-0011 is for research use only; downstream performance is not guaranteed.

Related topics covered:

column-free negative selection Untouched Human NK Cell (CD3-/CD56+ Enriched Fraction) isolation Untouched Human NK Cell (CD3-/CD56+ Enriched Fraction) purity verification Untouched Human NK Cell (CD3-/CD56+ Enriched Fraction) sorting protocol column-free isolation single-cell analysis (scRNA-seq) Untouched Human NK Cell (CD3-/CD56+ Enriched Fraction) isolation markers PBMC Untouched Human NK Cell (CD3-/CD56+ Enriched Fraction) sorting

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