RosetteSep™ HLA T Cell Enrichment Cocktail
Immunodensity negative selection cocktail
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RosetteSep™ HLA T Cell Enrichment Cocktail -
Label for RosetteSep™ HLA T Cell Enrichment Cocktail
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Required Products
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Density gradient medium for the isolation of mononuclear cells -
Dulbecco's Phosphate Buffered Saline with 2% Fetal Bovine Serum
Cell culture buffer -
Compatible antibodies for purity assessment of isolated cells
Overview
The RosetteSep™ HLA T Cell Enrichment Cocktail is designed to isolate T cells from whole blood by negative selection. Unwanted cells are targeted for removal with Tetrameric Antibody Complexes (TAC) recognizing non-T cells and red blood cells (RBCs). When centrifuged over a buoyant density medium such as RosetteSep™ DM-L (Catalog #15705) or Lymphoprep™ (Catalog #07801), the unwanted cells pellet along with the RBCs. The purified T cells are present as a highly enriched population at the interface between the plasma and the buoyant density medium. The desired cells are immediately ready for serology or flow cytometry crossmatch assays. This kit carries the CE mark and is available as a Class I in vitro diagnostic device in the European Union and Australia. To learn more about the regulatory status of this product in your specific region, contact info@stemcell.com.
Advantages:
• Fast and easy-to-use
• Requires no special equipment or training
• Isolated cells are untouched
• Requires no special equipment or training
• Isolated cells are untouched
Components:
- RosetteSep™ HLA T Cell Enrichment Cocktail (Catalog #15061HLA)
- RosetteSep™ HLA T Cell Enrichment Cocktail, 5 x 2 mL
- RosetteSep™ HLA T Cell Enrichment Cocktail (Catalog #15081HLA)
- RosetteSep™ HLA T Cell Enrichment Cocktail, 20 x 2 mL
Subtype:
Cell Isolation Kits
Cell Type:
T Cells
Species:
Human
Sample Source:
Buffy Coat; Whole Blood
Selection Method:
Negative
Application:
Cell Isolation; In Vitro Diagnostic
Brand:
RosetteSep
Area of Interest:
HLA; Immunology
Related Products
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Human Peripheral Blood Leukapheresis Pack, Fresh
Primary human cells, fresh -
Tube for density gradient centrifugation for in vitro diagnostic (IVD) applications -
EasySep™ Direct Human T Cell Isolation Kit
Immunomagnetic negative selection from whole blood kit
Labeling Antibodies
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Anti-Human CD3 Antibody, Clone UCHT1
Mouse monoclonal IgG1 antibody against human, chimpanzee CD3
Scientific Resources
Product Documentation
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Frequently Asked Questions
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.
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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.
Data and Publications
Data
Figure 1. Typical RosetteSep™ HLA T Cell Enrichment Profile
Starting with fresh whole blood the CD3+ cell content of the enriched fraction typically ranges from 90% - 97%. Red blood cells were removed by lysis prior to flow cytometry.
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.