EasySep® Human CD4+CD25+ T Cell Isolation Kit

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Immunomagnetic Positive Selection Kit

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  • EasySep® box
  • Human CD4+CD25high T Cell Isolation Kit Label
EasySep® box
The EasySep® Human CD4+CD25+ T Cell Isolation Kit is a simple two-step method designed to isolate CD4+CD25high cells from peripheral blood mononuclear cells (PBMC). CD4+ T cells are enriched from PBMC by negative selection using EasySep® Human CD4+ T Cell Enrichment Cocktail. The EasySep® cocktail contains a combination of Tetrameric Antibody Complexes (TAC) recognizing non-CD4+ T cells (CD8, CD14, CD16, CD19, CD20, CD36, CD56, CD66b, CD123, TCR¿/d) and glycophorin A on red blood cells. Labeled cells are separated using an EasySep® magnet without the use of columns. The enriched CD4+ T cells are then selected for CD25high cells by positive selection using EasySep® Human CD25 Positive Selection Cocktail. Cells are targeted with TAC recognizing CD25 and dextran-coated magnetic particles. Labeled cells are separated using an EasySep® magnet and remain in the tube while unwanted cells are poured off. The selection steps can be fully automated using RoboSep®.
Product Name Description Catalog # Size Price Quantity
EasySep® Human CD4+CD25 T Cell Isolation Kit Immunomagnetic positive selection of human CD4+CD25+ T cells 18062 For labeling up to 1 x 10e9 cells 824.00 USD      
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RoboSep Human CD4+CD25+ T Cell Isolation Kit with Filter Tips EasySep® kit with RoboSep® buffer and RoboSep® filter tips 18062RF For labeling up to 1 x 10e9 cells 855.00 USD      
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Order Offline:

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:
The isolation of CD4+CD25high T cells from PBMCs or leukopheresis
Components:
• EasySep® Human CD4+ T Cell Enrichment Cocktail
• EasySep® Magnetic Particles
• EasySep® Human CD25high Positive Selection Cocktail
• EasySep® Nanoparticles
• EasySep® Blocking Solution
Accessory Products:
Intended Use Statement: For Research Use Only. Not for Therapeutic or Diagnostic Use.
Equipment Required:
• "The Big Easy" EasySep® Magnet (Catalog #18001), or
• RoboSep® (Catalog #20000)
Product Type: Cell isolation products
Area of Interest: Immunology
Cell Isolation Product Type: Reagents
Cell Source: PBMC
Cell Type: Regulatory T cells, T cells
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 #
EasySep® Magnet Immunomagnetic Column-Free Magnet 18000
Easy 50 EasySep® Magnet Immunomagnetic Column-Free Magnet 18002
EasySep® Multistand Immunomagnetic Column-Free Multistand 18010
RoboSep™ RoboSep™ - The Fully Automated Cell Separator 20000
Complete Kit for Human CD4+CD25+ T Cells Immunomagnetic Positive Selection Kit 15862
Complete Kit for Human CD4+CD127lowCD25+ Regulatory T Cells Immunomagnetic Positive Selection Kit 15861
Complete Kit for Human CD4+CD127lowCD49d-CD25high Regulatory T Cells Immunomagnetic Positive Selection Kit 15864

FACS Profile Results With EasySep® Human CD4+CD25+ T Cell Isolation Kit


FACS Profile Results With EasySep® Human CD4+CD25+ T Cell Isolation Kit



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