The RosetteSep™ HLA Granulocyte Depletion Cocktail is designed to deplete granulocytes from whole blood. Unwanted cells are targeted for removal with Tetrameric Antibody Complexes recognizing CD66b and glycophorin A on red blood cells (RBCs). When centrifuged over a buoyant density medium such as Lymphoprep™ (a href="https://www.stemcell.com/products/lymphoprep.html">Catalog #18060), the unwanted cells pellet along with the RBCs. The desired cells are present as a highly enriched population depleted of granulocytes at the interface between the plasma and the buoyant density medium.
Please Note: In light of the new EU Regulation 2017/746 (IVDR) of the European Parliament and of the Council on in vitro diagnostic medical devices, which is expected to become effective in 2022, STEMCELL recently deregistered and removed the CE IVD claims associated with these products, and they are now labeled “For Research Use Only”. Please note there have been no changes to the form, function and quality assurances associated with these products. For more information please contact the Quality Assurance and Regulatory Department at qaschangenote@stemcell.com.
Subtype Cell Isolation Kits
Cell Type Granulocytes and Subsets
Species Human
Sample Source Buffy Coat, Whole Blood
Selection Method Depletion
Application Cell Isolation, In Vitro Diagnostic
Brand RosetteSep
Area of Interest HLA, Immunology
Data Figures
Figure 1. FACS Results Using RosetteSep™ HLA Granulocyte Depletion Kit
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.
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.
Thank you for your interest in IntestiCult™ Organoid Growth Medium (Human). Please provide us with your contact information and your local representative will contact you with a customized quote. Where appropriate, they
can also assist you with a(n):
Estimated delivery time for your area
Product sample or exclusive offer
In-lab demonstration
By submitting this form, you are providing your consent to STEMCELL Technologies Canada Inc. and its subsidiaries and affiliates (“STEMCELL”) to collect and use your information, and send you newsletters and emails in accordance with our
privacy policy. Please contact us with any questions that you may have. You can unsubscribe or change your email preferences at any time.
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.
