EasySep™ Human Th17 Cell Enrichment Kit

Immunomagnetic Positive Selection Kit

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From: 905 CAD

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Catalog # 18162
Immunomagnetic Positive Selection Kit
From: 905 CAD

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Overview

The EasySep™ Human Th17 Enrichment Kit is a two-step method designed to enrich human Th17 cells from fresh peripheral blood nucleated cells. CD4+CXCR3- T cells are first pre-enriched by negative selection using EasySep™ Human CD4+CXCR3- T Cell Pre-Enrichment Cocktail. Unwanted cells are targeted for removal with Tetrameric Antibody Complexes (TAC) recognizing CD8, CD14, CD16, CD19, CD20, CD36, CD56, CD66b, CD123, TCR/δ, glycophorin A, CD45RAhigh, CXCR3 and dextran-coated magnetic particles. The labeled cells are separated using an EasySep™ magnet without the use of columns. Desired cells are poured off into a new tube. Following pre-enrichment, CCR6 positive CD4+CXCR3- T cells are selected using EasySep™ Human CCR6 Positive Selection Cocktail. Cells are targeted with TAC recognizing CCR6 and dextran-coated magnetic particles. Labeled cells are separated using an EasySep™ magnet and remain in the tube while the unwanted cells are poured off.
Advantages:
• Fast and easy-to-use
• Up to 94% purity
• No columns required
Components:
  • EasySep™ Human Th17 Cell Enrichment Kit (Catalog #18162)
    • EasySep™ Human CD4+CXCR3- Pre-Enrichment Cocktail, 1 mL
    • EasySep™ Magnetic Particles, 2 x 1 mL
    • EasySep™ Human CCR6 Positive Selection Cocktail, 0.5 mL
    • EasySep™ Magnetic Nanoparticles, 1 mL
  • RoboSep™ Human Th17 Cell Enrichment Kit (Catalog #18162RF)
    • EasySep™ Human CD4+CXCR3- Pre-Enrichment Cocktail, 1 mL
    • EasySep™ Magnetic Particles, 2 x 1 mL
    • EasySep™ Human CCR6 Positive Selection Cocktail, 0.5 mL
    • EasySep™ Magnetic Nanoparticles, 1 mL
    • RoboSep™ Buffer (Catalog #20104) x 2
    • RoboSep™ Filter Tips (Catalog #20125) x 2
Magnet Compatibility:
• EasySep™ Magnet (Catalog #18000)
• “The Big Easy” EasySep™ Magnet (Catalog #18001)
• RoboSep™-S (Catalog #21000)
Subtype:
Cell Isolation Kits
Cell Type:
T Cells; T Cells, CD4+
Species:
Human
Sample Source:
Leukapheresis; PBMC
Selection Method:
Positive
Application:
Cell Isolation
Brand:
EasySep; RoboSep
Area of Interest:
Immunology

Technical Resources

Educational Materials

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Frequently Asked Questions

(16)

Can EasySep™ be used for either positive or negative selection?

Yes. The EasySep™ kits use either a negative selection approach by targeting and removing unwanted cells or a positive selection approach targeting desired cells. Depletion kits are also available for the removal of cells with a specific undesired marker (e.g. GlyA).

How does the separation work?

Magnetic particles are crosslinked to cells using Tetrameric Antibody Complexes (TAC). When placed in the EasySep™ Magnet, labeled cells migrate to the wall of the tube. The unlabeled cells are then poured off into a separate fraction.

Which columns do I use?

The EasySep™ procedure is column-free. That's right - no columns!

How can I analyze the purity of my enriched sample?

The Product Information Sheet provided with each EasySep™ kit contains detailed staining information.

Can EasySep™ separations be automated?

Yes. RoboSep™, the fully automated cell separator, automates all EasySep™ labeling and cell separation steps.

Can EasySep™ be used to isolate rare cells?

Yes. We recommend a cell concentration of 2x108 cells/mL and a minimum working volume of 100 µL. Samples containing 2x107 cells or fewer should be suspended in 100 µL of buffer.

Are the EasySep™ magnetic particles FACS-compatible?

Yes, the EasySep™ particles are flow cytometry-compatible, as they are very uniform in size and about 5000X smaller than other commercially available magnetic beads used with column-free systems.

Can the EasySep™ magnetic particles be removed after enrichment?

No, but due to the small size of these particles, they will not interfere with downstream applications.

Can I alter the separation time in the magnet?

Yes; however, this may impact the kit's performance. The provided EasySep™ protocols have already been optimized to balance purity, recovery and time spent on the isolation.

For positive selection, can I perform more than 3 separations to increase purity?

Yes, the purity of targeted cells will increase with additional rounds of separations; however, cell recovery will decrease.

How does the binding of the EasySep™ magnetic particle affect the cells? is the function of positively selected cells altered by the bound particles?

Hundreds of publications have used cells selected with EasySep™ positive selection kits for functional studies. Our in-house experiments also confirm that selected cells are not functionally altered by the EasySep™ magnetic particles.

If particle binding is a key concern, we offer two options for negative selection. The EasySep™ negative selection kits can isolate untouched cells with comparable purities, while RosetteSep™ can isolate untouched cells directly from whole blood without using particles or magnets.
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Product Applications

This product is designed for use in the following research area(s) as part of the highlighted workflow stage(s). Explore these workflows to learn more about the other products we offer to support each research area.

Data and Publications

Data

Start: 54% CD4+CXCR3-CCR6+ T Cells

Figure 1. Typical EasySep™ Human Th17 Cell Enrichment Profile

Starting with fresh PBMCs, the Th17 (CD4+CXCR3-CCR6+) cell content of the isolated fraction is typically 85 - 94%. In the above example, the purities of the start, pre-enriched, and final enriched fractions are 5.4%, 47.4%, and 91.4%, respectively. Intracellular staining for IL-17-producing cells typically ranges from 5 - 20% IL-17+ cells. These values vary widely among donors. IFN-γ-producing cells are typically < 5% of the enriched fraction.

STEMCELL TECHNOLOGIES INC.’S QUALITY MANAGEMENT SYSTEM IS CERTIFIED TO ISO 13485. PRODUCTS ARE FOR RESEARCH USE ONLY AND NOT INTENDED FOR HUMAN OR ANIMAL DIAGNOSTIC OR THERAPEUTIC USES UNLESS OTHERWISE STATED.