EasySep™ Human CD4+CD127lowCD49d- Regulatory T Cell Enrichment Kit

Immunomagnetic negative selection kit
Catalog #
19232_C
Immunomagnetic negative selection kit
From: 960 USD
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Required Products
  1. EasySep™ Magnet
    EasySep™ Magnet

    Magnet for column-free immunomagnetic separation

  2. EasySep™ Buffer
    EasySep™ Buffer

    Cell separation buffer

Overview

The EasySep™ Human CD4+CD127lowCD49d- Regulatory T Cell Enrichment Kit is designed to isolate CD4+CD127lowCD49d- cells from fresh peripheral blood mononuclear cells by negative selection. Unwanted cells are targeted for removal with Tetrameric Antibody Complexes recognizing non-CD4+ T cells, CD127high, CD49d+ cells and dextran-coated magnetic particles. The labeled cells are separated using the EasySep™ magnet without the use of columns. The desired cells are poured off into a new tube.
Advantages
• Fast, easy-to-use and column-free
• Up to 87.4% purity
• Isolated cells are untouched
Components
  • EasySep™ Human CD4+CD127lowCD49d- Regulatory T Cell Enrichment Kit (Catalog #19232)
    • EasySep™ Human CD4+CD127lowCD49d- Regulatory T Cell Enrichment Cocktail, 1 mL
    • EasySep™ Dextran Selection Cocktail, 1 mL
    • EasySep™ Magnetic Particles, 3 x 1 mL
  • RoboSep™ CD4+CD127lowCD49d- Regulatory T Cell Enrichment Kit (Catalog #19232RF)
    • EasySep™ Human CD4+CD127lowCD49d- Regulatory T Cell Enrichment Cocktail, 1 mL
    • EasySep™ Dextran Selection Cocktail, 1 mL
    • EasySep™ Magnetic Particles, 3 x 1 mL
    • RoboSep™ Buffer (Catalog #20104)
    • RoboSep™ Filter Tips (Catalog #20125)
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+, T Cells, Regulatory
Species
Human
Sample Source
PBMC
Selection Method
Negative
Application
Cell Isolation
Brand
EasySep, RoboSep
Area of Interest
Immunology

Scientific Resources

Product Documentation

Document Type Product Name Catalog # Lot # Language
Document Type
Product Information Sheet
Product Name
EasySep™ Human CD4+CD127lowCD49d- Regulatory T Cell Enrichment Kit
Catalog #
19232
Lot #
All
Language
English
Document Type
Product Information Sheet
Product Name
RoboSep™ CD4+CD127lowCD49d- Regulatory T Cell Enrichment Kit
Catalog #
19232RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Product Name
EasySep™ Human CD4+CD127lowCD49d- Regulatory T Cell Enrichment Kit
Catalog #
19232
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Product Name
EasySep™ Human CD4+CD127lowCD49d- Regulatory T Cell Enrichment Kit
Catalog #
19232
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Product Name
RoboSep™ CD4+CD127lowCD49d- Regulatory T Cell Enrichment Kit
Catalog #
19232RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Product Name
RoboSep™ CD4+CD127lowCD49d- Regulatory T Cell Enrichment Kit
Catalog #
19232RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 3
Product Name
RoboSep™ CD4+CD127lowCD49d- Regulatory T Cell Enrichment Kit
Catalog #
19232RF
Lot #
All
Language
English

Educational Materials (9)

Brochure
Tools For Your Immunology Research
Brochure
EasySep™ Cell Separation Technology
Wallchart
Regulatory T Cells
Wallchart
Human Immune Cytokines
Video
How to Isolate PBMCs from Whole Blood Using Density Gradient Centrifugation (Ficoll™ or Lymphoprep™)
1:37
How to Isolate PBMCs from Whole Blood Using Density Gradient Centrifugation (Ficoll™ or Lymphoprep™)
Video
How EasySep™ Magnetic Cell Separation Technology Works: Fast and Easy Cell Isolation
1:57
How EasySep™ Magnetic Cell Separation Technology Works: Fast and Easy Cell Isolation
Scientific Poster
Column-Free Isolation of Highly Purified and Functional Human Regulatory T Cells
Scientific Poster
Immunomagnetic Cell Isolation of Untouched Human Regulatory T Cells
Scientific Poster
Rapid Isolation of Highly Purified, Functional and Expandable Human Regulatory T Cells

Frequently Asked Question

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.
Read More

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

Typical EasySep™ Human CD4+CD127lowCD49d- Regulatory T Cell Enrichment Profile

Figure 1. Typical EasySep™ Human CD4+CD127lowCD49d- Regulatory T Cell Enrichment Profile

Starting with fresh PBMCs, the CD3+CD4+CD127lowCD49d-CD25+FOXP3+ cell content of the enriched fraction typically ranges from 57.4 - 87.4%. Donor to donor variability is unavoidable and may affect the purity of the enriched fraction. In the example above, the purities of the start and enriched fractions are 1.84% and 84.1%, respectively.

Publications (1)

The Journal of Immunology 2015 A Transendocytosis Model of CTLA-4 Function Predicts Its Suppressive Behavior on Regulatory T Cells Hou TZ et al.

Abstract

Manipulation of the CD28/CTLA-4 pathway is at the heart of a number of immunomodulatory approaches used in both autoimmunity and cancer. Although it is clear that CTLA-4 is a critical regulator of T cell responses, the immunological contexts in which CTLA-4 controls immune responses are not well defined. In this study, we show that whereas CD80/CD86-dependent activation of resting human T cells caused extensive T cell proliferation and robust CTLA-4 expression, in this context CTLA-4 blocking Abs had no impact on the response. In contrast, in settings where CTLA-4(+) cells were present as regulators�
View All Publications

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