What starting material and cell density are specified?

Use fresh or frozen human PBMCs prepared as a uniform single-cell suspension. The documented setup adjusts cells to 1×108 cells/mL and uses 100 µL, or 1×107 cells, per assay. Pass the suspension through a 70 µm strainer before separation.

CS-0018

Human Naïve T Cell Sorting Kit

Column-Free Negative Human

Cell Sorts™ Human Naïve T Cell Sorting Kit (CS-0018) enriches an untouched naïve pan T-cell fraction from fresh or cryopreserved human PBMCs through column-free negative selection. A biotinylated isolation cocktail labels non-target cells, and Streptavidin Nanobeads retain the labeled fraction on a compatible tube magnet. The recovered suspension contains target-enriched cells that are not directly labeled by the isolation antibodies or beads.

Each documented assay starts with 1×10⁷ cells and uses 40 µL cocktail plus 30 µL pre-washed streptavidin beads. One representative healthy-donor analysis reported 95.2% CD3+CD45RA+CD45RO-CD197+ cells after sorting; this is not a guaranteed specification. The workflow accepts fresh or frozen PBMCs, includes a 300 × g wash, and requires a 5-minute magnetic hold. Verify phenotype, purity, viability, and recovery before downstream use.

Sorting Purity

Representative result 95.2%

Sorting Time

At least 25 min documented timed steps after PBMC preparation

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Product Features of the Human Naïve T Cell Sorting Kit

CS-0018 uses a documentation-aligned column-free negative-selection workflow to enrich an untouched human naïve pan T-cell fraction from fresh or frozen PBMCs, with defined assay dosing, a 300 × g wash, pre-washed streptavidin beads, and tube-based magnetic retention.

Column-free negative selection - Labeled non-target cells are magnetically retained in a tube without a separation column

Untouched target-enriched fraction - The recovered naïve pan T-cell fraction is not directly labeled by the isolation antibodies or streptavidin beads

Fresh and frozen PBMC compatibility - The documented frozen-sample workflow includes 100 µg/mL DNase I treatment for at least 15 minutes before washing

Defined assay dosing - One assay processes 1×10⁷ cells with 40 µL isolation cocktail and 30 µL pre-washed streptavidin beads

Evidence-qualified phenotype - One representative healthy-donor analysis reported 95.2% CD3+CD45RA+CD45RO-CD197+ cells after separation; results are not guaranteed

Flexible pack sizes - Documented 10-, 100-, and 200-assay formats support experimental planning across different study scales

CS-0018 uses column-free negative selection to enrich an untouched human naïve pan T-cell fraction from fresh or frozen PBMCs. The supplied document does not state a validated total processing time; the listed post-preparation timed steps total at least 25 minutes, with mixing, transfers, counting, optional collection, and QC additional.

How the Human Naïve T Cell Sorting Kit Works

Four stages summarizing the documented fresh-or-frozen PBMC naïve pan T-cell workflow

Step 1: Prepare Fresh or Frozen PBMCs

Prepare fresh PBMCs by density-gradient centrifugation or treat thawed PBMCs with 100 µg/mL DNase I for at least 15 minutes at room temperature. Wash, pass through a 70 µm strainer, count, and adjust to 1×10⁸ cells/mL.

Step 2: Label Non-Target Cells

Transfer 100 µL, or 1×10⁷ cells, to a 2 mL tube. Add 40 µL Human Naïve Pan T Cell Isolation Cocktail, mix by pipetting 6-8 times, and incubate for 5 minutes at room temperature.

Step 3: Wash Cells and Prepare Beads

Bring the cell suspension to 2 mL with separation buffer, centrifuge at 300 × g for 5 minutes, discard the supernatant, and resuspend in 100 µL. Separately wash 30 µL streptavidin beads in 1 mL buffer on the magnet for 5 minutes.

Step 4: Bind, Capture, and Collect

Resuspend the washed beads with the 100 µL cell suspension, mix 6-8 times, and incubate for 5 minutes at room temperature. Add buffer to 2.5 mL, mix thoroughly, place on the magnet for 5 minutes, and transfer the unretained naïve pan T-cell-enriched fraction to a clean tube.

Applications for the Human Naïve T Cell Sorting Kit

Potential downstream uses for the enriched untouched naïve pan T-cell fraction are listed below. Confirm CD3, CD45RA, CD45RO, and CD197/CCR7 phenotype, subset composition, purity, viability, recovery, and functional suitability.

Flow Cytometric Quality Control - Confirm CD3, CD45RA, CD45RO, and CD197/CCR7 phenotype with a laboratory-validated gating strategy Naïve-versus-Memory T-Cell Phenotyping - Characterize the enriched fraction while monitoring residual memory and non-T-cell populations Naïve CD4 and CD8 Composition Analysis - Quantify helper and cytotoxic subsets within the recovered pan T-cell fraction Controlled T-Cell Activation and Differentiation - Establish fit-for-purpose stimulation and lineage-differentiation experiments after post-sort QC Primary Immune-Response Research - Study activation, clonal expansion, and differentiation from a qualified naïve T-cell starting population TCR Repertoire and Molecular Profiling - Prepare DNA, RNA, or protein inputs when assay-specific acceptance criteria are met Single-Cell Analysis - Prepare qualified cell suspensions for single-cell workflows after confirming viability, recovery, and platform compatibility PBMC Preparation and Separation Development - Optimize fresh-versus-frozen sample handling, DNase treatment, reagent scaling, magnetic capture, and recovery

The listed applications are potential research uses, not guaranteed performance claims. Confirm naïve pan T-cell identity, subset composition, purity, viability, recovery, and assay-specific suitability.

Kit Components

CS-0018 includes the documented Human Naïve Pan T Cell Isolation Cocktail, Human Naïve Pan T Beads Streptavidin 1.0-N, and user manual. Separation buffer, a compatible tube magnet, sterile tubes, PBMC-preparation materials, a centrifuge, a 70 µm strainer, and post-sort QC reagents are required separately.

Human Naïve Pan T Cell Isolation Cocktail
Human Naïve Pan T Beads Streptavidin 1.0-N
User manual

Storage & Shelf Life

Storage

2-8°C protected from light; avoid freezing and freeze-thaw cycles

Shelf Life

12 months

Human Naïve T Cell Sorting Kit — Frequently Asked Questions

Everything you need to know about the CS-0018 Human Naïve T Cell Sorting Kit

Click each question to expand the answer

CS-0018 enriches an untouched human naïve pan T-cell fraction from fresh or cryopreserved PBMCs. The supplied document defines the representative target phenotype as CD3+CD45RA+CD45RO-CD197+; laboratories should use a fit-for-purpose panel and gating strategy to confirm identity and residual populations.

The kit uses column-free negative selection. A biotinylated isolation cocktail labels non-target cells, pre-washed streptavidin beads bind the labeled fraction, and a compatible tube magnet retains those cells while the unretained target-enriched suspension is transferred to a fresh tube.

No. In the documented negative-selection design, antibodies and streptavidin beads are used to label and retain non-target cells. The recovered target-enriched fraction is not directly labeled by the isolation reagents, but identity, purity, viability, recovery, and functional suitability should still be confirmed.

In one representative healthy-donor PBMC experiment, CD3+CD45RA+CD45RO-CD197+ cells increased from 6.0% before separation to 95.2% afterward. This is a representative example rather than a guaranteed specification; donor, sample quality, handling, magnet performance, and gating can affect results.

The document does not state a validated total processing time. After PBMC preparation, the listed timed steps total at least 25 minutes: 5-minute cocktail incubation, 5-minute cell wash centrifugation, 5-minute bead pre-wash on the magnet, 5-minute bead incubation, and 5-minute final magnetic hold. Mixing, transfers, counting, optional collection, and QC require additional time.

Use fresh or frozen human PBMCs prepared as a uniform single-cell suspension. The documented setup adjusts cells to 1×10⁸ cells/mL and uses 100 µL, or 1×10⁷ cells, per assay. Pass the suspension through a 70 µm strainer before separation.

Before sorting, incubate thawed PBMCs with DNase I at a final concentration of 100 µg/mL for at least 15 minutes at room temperature. Then wash twice at 300 × g for 5 minutes, filter through a 70 µm strainer, count, and adjust the cell concentration.

For 1×10⁷ cells, use 40 µL isolation cocktail and 30 µL pre-washed streptavidin beads. Scale proportionally above 1×10⁷ cells. For lower inputs, the document retains the same minimum reagent volumes. A 5 mL flow tube is listed for up to 1×10⁸ cells, while a 10 or 15 mL tube is listed for up to 4×10⁸ cells.

Use a laboratory-validated flow-cytometry panel that can assess CD3, CD45RA, CD45RO, and CD197/CCR7, together with viability and singlet gating. Report purity, viability, recovery, and residual populations for the actual donor and preparation rather than relying on the representative result.

The documented components are Human Naïve Pan T Cell Isolation Cocktail, Human Naïve Pan T Beads Streptavidin 1.0-N, and the user manual. Store the kit at 2-8°C for up to 12 months, protected from light, and avoid freezing or freeze-thaw cycles. A compatible magnet, separation buffer, tubes, centrifuge, and QC reagents are required separately.