EasySep™ Mouse ILC2 Enrichment Kit

Immunomagnetic negative selection kit

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EasySep™ Mouse ILC2 Enrichment Kit

Immunomagnetic enrichment of untouched mouse ILC2 cells

1 x 109 cells
Catalog #19842
687 USD

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Overview Related Products Scientific Resources Product Applications Data and Publications

Overview

This product is designed to enrich ILC2's (Group 2 Innate Lymphoid Cells) from single-cell suspensions of lung tissue or other tissues by negative selection. Unwanted cells are targeted for removal with biotinylated antibodies directed against non-ILC2 cells and streptavidin-coated magnetic particles (RapidSpheres™). Labeled cells are separated using an EasySep™ magnet without the use of columns.
Advantages:
• Fast, easy-to-use and column-free
• Isolated cells are untouched
• Facilitates rapid flow sorting of ILC2s
Components:
  • EasySep™ Mouse ILC2 Enrichment Kit (Catalog #19842)
    • EasySep™ Mouse Enrichment Cocktail, 0.5 mL
    • EasySep™ Streptavidin RapidSpheres™ 50001, 1.0 mL
Magnet Compatibility:
EasySep™ Magnet (Catalog #18000)
Subtype:
Cell Isolation Kits
Cell Type:
Innate Lymphoid Cells
Species:
Mouse
Sample Source:
Other
Selection Method:
Negative
Application:
Cell Isolation
Brand:
EasySep
Area of Interest:
Immunology

Scientific Resources

Product Documentation

Educational Materials

(9)
Brochure
Tools For Your Immunology Research
Brochure
Cell Enrichment Prior to Cell Sorting
Brochure
EasySep™ Cell Separation Technology
Technical Bulletin
Gating Strategy for the Identification of ILC2s
Wallchart
Human Immune Cytokines
Video
3:58
Comparison of Mouse Immune Cell Isolation Protocols: EasySep™ vs. Column-Based Magnetic Separation
Video
1:57
How EasySep™ Magnetic Cell Separation Technology Works: Fast and Easy Cell Isolation
Scientific Poster
Releasable RapidSpheres Enable Immunomagnetic Purification of Highly Viable and Functional Immune Cells from Complex Tissues in Less Than 30 Minutes
Scientific Poster
Magnetic Enrichment of Mouse ILC2s from the Lung

Frequently Asked Questions

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.

Research Area Workflow Stages for
Workflow Stages
Innate Lymphoid Cell Research
Cell Sourcing
Cell Isolation
Cell Culture and Characterization

Data and Publications

Data

Starting with a naïve mouse lung single-cell suspension, the ILC2 content (CD45+Lin-CD278+CD90.2+ST2+) of the final enriched fraction typically ranges from 2.2 - 7.1%. In the above example, the percentage of ILC2s in the start and final enriched fractions are 0.8% and 6.5% (or 0.9% and 22.3% of CD45+ cells), respectively. NOTE: The ILC2 content of the start fraction typically ranges from 0.1 - 1%.

Publications

(2)
Current protocols in immunology 2019 jun

Identification of Group 2 Innate Lymphoid Cells in Mouse Lung, Liver, Small Intestine, Bone Marrow, and Mediastinal and Mesenteric Lymph Nodes.

M. Romera-Hern\'andez et al.
Abstract
View Publication

Abstract

Innate lymphoid cells (ILCs) are a heterogeneous family of lymphocytes that populate barrier and non-barrier tissues. ILCs regulate immune responses to pathogens and commensals but also sustain metabolic homeostasis, tissue remodeling after injury and establish dialogue with the nervous system. ILCs rapidly become activated in the absence of adaptive antigen receptors by responding to signaling molecules provided by hematopoietic or non-hematopoietic cells. Here we provide protocols designed for processing the lung, liver, small intestine, bone marrow, mediastinal and mesenteric lymph nodes in order to obtain a purified leukocyte fraction of cells, in which ILC2 enrichment is optimized. In addition, we describe in detail the methodologies used to activate ILC2s and the assays necessary for the detection of their effector cytokines. We highlight the differences in ILC2 characterization within distinct tissues that we have recently identified. {\textcopyright} 2019 by John Wiley Sons, Inc.
Immunity 2016 JUL

Allergen-Experienced Group 2 Innate Lymphoid Cells Acquire Memory-like Properties and Enhance Allergic Lung Inflammation.

Martinez-Gonzalez I et al.
Abstract
View Publication

Abstract

Group 2 innate lymphoid cells (ILC2s) in the lung are stimulated by inhaled allergens. ILC2s do not directly recognize allergens but they are stimulated by cytokines including interleukin (IL)-33 released by damaged epithelium. In response to allergens, lung ILC2s produce T helper 2 cell type cytokines inducing T cell-independent allergic lung inflammation. Here we examined the fate of lung ILC2s upon allergen challenges. ILC2s proliferated and secreted cytokines upon initial stimulation with allergen or IL-33, and this phase was followed by a contraction phase as cytokine production ceased. Some ILC2s persisted long after the resolution of the inflammation as allergen-experienced ILC2s and responded to unrelated allergens more potently than naive ILC2s, mediating severe allergic inflammation. The allergen-experienced ILC2s exhibited a gene expression profile similar to that of memory T cells. The memory-like properties of allergen-experienced ILC2s may explain why asthma patients are often sensitized to multiple allergens.
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