Human CD4+ T Cell Sorting Kit CS-0006 for column-based T cell isolation | Cell Sorts™
CS-0006

Human CD4+ T Cell Sorting Kit

Column-Based Negative Human

Cell Sorts™ Human CD4+ T Cell Sorting Kit (CS-0006) enriches untouched CD4+ T cells from human PBMCs using column-based negative selection. A depletion cocktail and magnetic nanobeads label non-target cells, which are retained in a compatible L column; the unlabeled CD4+ T-cell fraction is recovered in the flow-through.

Supplied technical documentation reports purity above 90%, with results up to 99% under tested conditions. The protocol scales reagents to cell input and supports up to 1×10⁸ cells per L column. Recovered cells can be evaluated for flow cytometry, culture, activation, cytokine, polarization, and single-cell workflows after laboratory-specific purity, viability, recovery, and yield checks. For research use only.

Sorting Purity
Above 90%, up to 99%
Sorting Time
Protocol-defined steps

Product Features of the Human CD4+ T Cell Sorting Kit

CS-0006 uses a documented column-based negative-selection workflow to enrich untouched human CD4+ T cells from Ficoll-prepared PBMCs.

Column-based negative selection — A depletion cocktail and magnetic nanobeads label non-target cells for retention in a compatible L column
Untouched CD4+ T-cell fraction — Target cells are collected in the flow-through without direct antibody or bead labeling
Documented purity above 90%, up to 99% — Performance remains sample- and process-dependent
Defined reagent scaling — Use 10 µL isolation cocktail and 10 µL nanobeads per 1×10⁷ starting cells
Controlled column loading — Use at least 500 µL sample volume and no more than 1×10⁸ total cells per L column
Workflow-focused quality control — Measure post-sort purity, viability, recovery, and total yield for each laboratory workflow

CS-0006 uses column-based negative selection to enrich untouched human CD4+ T cells from Ficoll-prepared PBMCs. The technical documentation defines the individual incubation, centrifugation, column-equilibration, loading, and wash steps but does not state a validated total processing time.

How the Human CD4+ T Cell Sorting Kit Works

Simple 4-step protocol for rapid cell isolation

1
CS-0006 step 1: prepare and count a Ficoll-separated human PBMC suspension for CD4+ T-cell enrichment

Step 1: Prepare the PBMC Suspension

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

2
CS-0006 step 2: label non-target cells, wash, and centrifuge at 400×g for 5 minutes

Step 2: Label and Wash Non-Target Cells

Add 10 µL isolation cocktail per 1×10⁷ cells and incubate for 10 minutes at room temperature. Add 10× buffer volume, centrifuge at 400×g for 5 minutes, discard the supernatant, and restore the original volume.

3
CS-0006 step 3: bind magnetic nanobeads and equilibrate the compatible L column with 3 mL buffer

Step 3: Bind Nanobeads and Prepare the Column

Mix the nanobeads for 5–30 seconds, add 10 µL per 1×10⁷ cells, and incubate for 5 minutes at room temperature. In parallel, equilibrate a compatible L column with 3 mL FACS buffer.

4
CS-0006 step 4: load at least 500 µL, collect untouched CD4+ T cells in the flow-through, and wash the column

Step 4: Load, Separate, and Collect

Adjust the sample to at least 500 µL and do not exceed 1×10⁸ cells per L column. Load by gravity, collect the CD4+ T-cell flow-through, then wash the column with 3 mL buffer and combine the fractions as directed.

Applications for the Human CD4+ T Cell Sorting Kit

This Human CD4+ T Cell Sorting Kit supports a wide range of downstream applications for CD4+ T Cell isolation and column-based negative selection workflows from human PBMCs.

CD4+ T Cell Phenotyping — Prepare enriched populations for multiparameter flow-cytometry panels after purity and viability assessment Activation and Proliferation Studies — Evaluate receptor-driven responses in untouched CD4+ T-cell preparations Helper T Cell Differentiation — Support laboratory-validated Th1, Th2, Th17, or regulatory T-cell polarization workflows Cytokine Analysis — Study cytokine secretion and signaling after appropriate stimulation and controls Cell Culture — Establish downstream expansion or functional-assay workflows using the recovered flow-through fraction Immunology and Host-Response Research — Investigate CD4+ T-cell biology in infectious-disease, inflammatory, or immune-regulation models Single-Cell and Molecular Analysis — Prepare samples for RNA, TCR, or single-cell workflows when input-quality requirements are met Drug Discovery — Assess candidate immunomodulators in laboratory-validated CD4+ T-cell assays

In addition, the Human CD4+ T 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 CD4+ T Cell Sorting Kit contains everything needed for high-purity column-based negative selection from human PBMCs.

  • Human CD4+ T cell Isolation Cocktail
  • Ultra-Nanobeads

Storage & Shelf Life

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

Human CD4+ T Cell Sorting Kit — Frequently Asked Questions

Everything you need to know about the CS-0006 Human CD4+ T Cell Sorting Kit

Click each question to expand the answer

CS-0006 is intended for enriching untouched human CD4+ T cells from PBMCs prepared by Ficoll density-gradient separation. The supplied instructions advise against using a red-blood-cell-lysis preparation as the starting sample for this workflow.
An isolation cocktail labels non-target cells, and magnetic nanobeads bind the labeled population. A compatible magnetized L column retains those cells while the untouched CD4+ T-cell fraction is collected in the flow-through and subsequent wash.
No. The depletion reagents are directed to non-target cells. CD4+ T cells are recovered without direct antibody or bead labeling, although laboratories should confirm the phenotype and functional suitability required for each downstream assay.
The supplied technical documentation reports purity above 90% and up to 99% under tested conditions. One representative example increased the CD4+ fraction from 25.3% to 94.8%. Results vary with donor biology, PBMC quality, starting composition, and protocol execution.
No validated total processing time is stated in the supplied documentation. It specifies a 10-minute cocktail incubation, a 5-minute nanobead incubation, a 5-minute centrifugation, and the required column-equilibration, loading, and wash steps.
For each 1×10⁷ starting cells, resuspend in 100 µL buffer, add 10 µL isolation cocktail, and later add 10 µL well-mixed nanobeads. Scale all volumes proportionally and follow the current instructions for use.
Use a minimum sample loading volume of 500 µL and do not exceed 1×10⁸ total cells per L column. Use additional columns for larger samples. Equilibrate each column with 3 mL buffer before loading.
The documented workflow uses FACS buffer prepared as 1× PBS with 2% FBS and 0.02% EDTA, a compatible L column, and a compatible magnetic column separator. Routine cell-counting and centrifugation equipment are also required.
The supplied instructions describe a self-provided Fc-receptor blocker as optional. It may help reduce nonspecific labeling and support purity or viability, particularly with challenging PBMC preparations. Follow the blocker manufacturer’s instructions and validate its effect in your workflow.
Review PBMC preparation and viability, aggregation, cell counting, reagent scaling, nanobead mixing, incubation conditions, the 400×g wash step, column equilibration, loading volume, column capacity, gravity flow, and collection of both the initial flow-through and 3 mL wash. Confirm purity, viability, recovery, and yield by an appropriate assay.

Related topics covered:

column-based negative selection CD4+ T Cell isolation CD4+ T Cell purity verification CD4+ T Cell sorting protocol column-based vs column-free single-cell analysis (scRNA-seq) CD4+ T Cell isolation markers PBMC CD4+ T Cell sorting

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