EasySep™ Human Th17 Cell Enrichment Kit

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Immunomagnetic Positive Selection Kit

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  • Box top label for EasySep Human Th17 Cell Enrichment Kit For labeling 10e9 cells
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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¿/d, 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. The selection steps can be fully automated using RoboSep™.
Product Name Description Catalog # Size Price Quantity
EasySep™ Human Th17 Cell Enrichment Kit Immunomagnetic positive selection of human Th17+ cells 18162 For labeling up to 1 x 10e9 cells 790.00 USD      
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RoboSep™ Human Th17 Cell Enrichment Kit EasySep™ kit with RoboSep™ buffer and RoboSep™ filter tips 18162RF For labeling up to 1 x 10e9 cells 858.00 USD      
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Call us: +1.604.877.0713  
Toll Free: 1.800.667.0322  
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Fax us: +1.604.877.0704
Toll Free Fax: 1.800.567.2899

Recommended for:
Enrichment of Th17 cells from freshly isolated peripheral blood mononuclear cells, leukapheresis or buffy coat less than 24 hours old.
Components:
• EasySep™ Human CD4+CXCR3- Pre-Enrichment Cocktail, 1.0 mL
• EasySep™ Magnetic Particles, 2 x 1.0 mL
• EasySep™ Human CCR6 Positive Selection Cocktail, 0.5 mL
• EasySep™ Magnetic Nanoparticles, 1.0 mL
Intended Use Statement: For Research Use Only. Not for Therapeutic or Diagnostic Use.
Equipment Required:
• EasySep™ Magnet (Catalog #18000), or
• “The Big Easy” EasySep™ Magnet (Catalog #18001), or
• RoboSep™ (Catalog #20000)
Advantages:
This kit provides a method to obtain live, non-activated Th17 cells that produce high levels of IL-17 and minimal amounts of IFN-g. Enriched cells could be used immediately for ex vivo studies or for further expansion.
 
Product Type: Cell isolation products
Area of Interest: Immunology
Cell Isolation Product Type: Reagents
Cell Source: Leukapheresis, PBMC
Cell Type: Other T cell subsets
Popular Product Line: EasySep, RoboSep
Selection Method: Positive
Species: Human

Procedures and instruction manuals:

Educational resources:

MSDS:

FAQS:


  • HOW EASYSEP™ WORKS

    • Q. CAN EASYSEP™ BE USED FOR EITHER POSITIVE OR NEGATIVE SELECTION?
      A. 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).
    • Q. HOW DOES THE SEPARATION WORK?
      A. 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.
    • Q. WHICH COLUMNS DO I USE?
      A. The EasySep™ procedure is column-free. That's right - no columns!
    • Q. HOW CAN I ANALYZE THE PURITY OF MY ENRICHED SAMPLE?
      A. The Product Information Sheet provided with each EasySep™ kit contains detailed staining information.
    • Q. CAN EASYSEP™ SEPARATIONS BE AUTOMATED?
      A. Yes. RoboSep™, the fully automated cell separator, automates all EasySep™ labeling and cell separation steps.
    • Q. CAN EASYSEP™ BE USED TO ISOLATE RARE CELLS?
      A. 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.

    EASYSEP™ MAGNETIC PARTICLES

    • Q. ARE THE EASYSEP™ MAGNETIC PARTICLES FACS-COMPATIBLE?
      A. 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.
    • Q. CAN THE EASYSEP™ MAGNETIC PARTICLES BE REMOVED AFTER ENRICHMENT?
      A. No, but due to the small size of these particles, they will not interfere with downstream applications.
    • Q. HOW DOES THE BINDING OF THE EASYSEP™ MAGNETIC PARTICLE AFFECT THE CELLS? IS THE FUNCTION OF POSITIVELY SELECTED CELLS ALTERED BY THE BOUND PARTICLES?
      A. 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.

    MODIFYING THE EASYSEP™ PROTOCOL

    • Q. CAN I ALTER THE SEPARATION TIME IN THE MAGNET?
      A. 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.
    • Q. FOR POSITIVE SELECTION, CAN I PERFORM MORE THAN 3 SEPARATIONS TO INCREASE PURITY?
      A. Yes, the purity of targeted cells will increase with additional rounds of separations; however, cell recovery will decrease.

Product Name

Description

Catalog #
RoboSep™ Buffer RoboSep™ Buffer 20104
The Big Easy EasySep® Magnet Immunomagnetic Column-Free Magnet 18001
RoboSep™ RoboSep™ - The Fully Automated Cell Separator 20000
EasySep® Magnet Immunomagnetic Column-Free Magnet 18000

Start: 5.4% CD4+CXCR3-CCR6+ T Cells


Start: 5.4% CD4+CXCR3-CCR6+ T Cells



Pre-enriched: 47.4% CD4+CXCR3-CCR6+ T Cells


Pre-enriched: 47.4% CD4+CXCR3-CCR6+ T Cells



Enriched: 91.4% CD4+CXCR3-CCR6+ T Cells of Pre-enriched Fraction


Enriched: 91.4% CD4+CXCR3-CCR6+ T Cells of Pre-enriched Fraction
Starting with fresh peripheral blood nucleated cells, the CD4+CXCR3-CCR6+ T cell content of the enriched fraction typically ranges from 85 - 94%. Intracellular staining for IL-17 producing cells typically ranges from 5 - 20% IL-17+ cells. These values vary widely amongst donors. IFN-γ producing cells are typically <5% of the enriched fraction.


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