RosetteSep™ Human ILC2 Enrichment Kit
Immunodensity negative selection cocktail
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Required Products
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Lymphoprep™Density gradient medium for the isolation of mononuclear cells
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Dulbecco's Phosphate Buffered Saline with 2% Fetal Bovine SerumCell culture buffer
Overview
The RosetteSep™ Human ILC2 Enrichment Cocktail is designed to enrich human ILC2s from whole blood by negative selection. Unwanted cells are targeted for removal with Tetrameric Antibody Complexes recognizing non-ILC2s and glycophorin A on red blood cells (RBCs). When centrifuged over a density gradient medium such as Lymphoprep™ (Catalog #07801), the unwanted cells pellet along with the RBCs. The ILC2 cells are present as a highly enriched population at the interface between the plasma and the density gradient medium.
Advantages
• Fast and easy-to-use
• Requires no special equipment or training
• Untouched, viable cells
• Can be combined with SepMate™ for consistent, high-throughput sample processing
• Facilitates rapid flow sorting of ILC2s
• Requires no special equipment or training
• Untouched, viable cells
• Can be combined with SepMate™ for consistent, high-throughput sample processing
• Facilitates rapid flow sorting of ILC2s
Components
- RosetteSep™ Human ILC2 Enrichment Kit (Catalog #15382)
- RosetteSep™ Human ILC2 Enrichment Cocktail, 5 x 2 mL
Subtype
Cell Isolation Kits
Cell Type
Innate Lymphoid Cells
Species
Human
Sample Source
Whole Blood
Selection Method
Negative
Brand
RosetteSep
Area of Interest
Immunology
Scientific Resources
Product Documentation
| Document Type | Product Name | Catalog # | Lot # | Language |
|---|---|---|---|---|
|
Document Type
Product Information Sheet
|
Product Name
RosetteSep™ Human ILC2 Enrichment Kit
|
Catalog #
15382 |
Lot #
All |
Language
English |
|
Document Type
Safety Data Sheet
|
Product Name
RosetteSep™ Human ILC2 Enrichment Kit
|
Catalog #
15382 |
Lot #
All |
Language
English |
Educational Materials (7)
Brochure
Cell Enrichment Prior to Cell Sorting
Technical Bulletin
Gating Strategy for the Identification of ILC2s
Wallchart
Innate Lymphoid Cells
Wallchart
Human Immune Cytokines
Video
1:46
How to Isolate PBMCs from Whole Blood Using the SepMate™ PBMC Isolation Tubes: 15-Minute Protocol
Video
2:19
Isolate Cells from Whole Blood without Columns or Magnets: RosetteSep™ Immunodensity Cell Separation
Scientific Poster
Fast and Efficient Enrichment of Functional ILC2 From Human Whole Blood
Frequently Asked Question
What is RosetteSep™?
RosetteSep™ is a rapid cell separation procedure for the isolation of purified cells directly from whole blood, without columns or magnets.
How does RosetteSep™ work?
The antibody cocktail crosslinks unwanted cells to red blood cells (RBCs), forming rosettes. The unwanted cells then pellet with the free RBCs when centrifuged over a density centrifugation medium (e.g. Ficoll-Paque™ PLUS, Lymphoprep™).
What factors affect cell recovery?
The temperature of the reagents can affect cell recovery. All reagents should be at room temperature (sample, density centrifugation medium, PBS, centrifuge) before performing the isolations. Layering can also affect recovery so be sure to carefully layer the sample to avoid mixing with the density centrifugation medium as much as possible. Be sure to collect the entire enriched culture without disturbing the RBC pellet. A small amount of density centrifugation medium can be collected without worry.
Which cell samples can RosetteSep™ be used with?
RosetteSep™ can be used with leukapheresis samples, bone marrow or buffy coat, as long as: the concentration of cells does not exceed 5 x 107 per mL (can dilute if necessary); and there are at least 100 RBCs for every nucleated cell (RBCs can be added if necessary).
Can RosetteSep™ be used with previously frozen or cultured cells?
Yes. Cells should be re-suspended at 2 - 5 x 107 cells / mL in PBS + 2% FBS. Fresh whole blood should be added at 250 µL per mL of sample, as a source of red cells.
Can RosetteSep™ be used to enrich progenitors from cord blood?
Yes. Sometimes cord blood contains immature nucleated red cells that have a lower density than mature RBCs. These immature red cells do not pellet over Ficoll™, which can lead to a higher RBC contamination than peripheral blood separations.
Does RosetteSep™ work with mouse cells?
No, but we have developed EasySep™, a magnetic-based cell isolation system which works with mouse and other non-human species.
Which anticoagulant should be used with RosetteSep™?
Peripheral blood should be collected in heparinized Vacutainers. Cord blood should be collected in ACD.
Should the anticoagulant be washed off before using RosetteSep™?
No, the antibody cocktail can be added directly to the sample.
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.
Research Area
Workflow Stages
Workflow Stages for
Innate Lymphoid Cell Research
Data and Publications
Data
Starting with fresh whole blood, the ILC2 content (Lin-CD45+CD294+CD127+CD161+) of the enriched fraction typically ranges from 0.44 - 53%. In the above example, the percentage of ILC2s in the start and final enriched fractions are 0.02% and 10.1%, respectively.
Quality Statement:
PRODUCTS ARE FOR RESEARCH USE ONLY AND NOT INTENDED FOR HUMAN OR ANIMAL DIAGNOSTIC OR THERAPEUTIC USES UNLESS OTHERWISE STATED. FOR ADDITIONAL INFORMATION ON QUALITY AT STEMCELL, REFER TO WWW.STEMCELL.COM/COMPLIANCE.
