RosetteSep™ DM-L is a density gradient medium designed specifically for use with RosetteSep™ cocktails for the enrichment of specific human lymphocyte populations (CD3+ T cells, CD4+ T cells, CD8+ T cells, and B cells) from whole blood. RosetteSep™ DM-L has a density of 1.081 g/mL.
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.
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.
Data Figures
Figure 1. Minimal Granulocyte Contamination Using RosetteSep™ DM-L Density Medium
Granulocyte contamination is typically less than 1% when enriching lymphocytes from fresh whole blood using RosetteSep™. There was no significant difference in the level of granulocyte contamination when using RosetteSep™ DM-L in place of Ficoll-paque PLUS to enrich lymphocytes (N=11)
Figure 2. Rosettesep™ DM-L Improves the Recovery of Lymphocytes Enriched with Rosettesep™ from Fresh Blood
Figure 3. Cell Purity Is Equivalent or Higher for Rosettesep™ Lymphocyte Enrichment Using Rosettesep™ DM-L Instead of Ficoll-paque®
Protocols and Documentation
Find supporting information and directions for use in the Product Information Sheet or explore additional protocols below.
Document Type
Product Name
Catalog #
Lot #
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Document Type
Product Information Sheet 1
Product Name
RosetteSep™ DM-L Density Medium
Catalog #
15705
Lot #
Lots above 1000053496
Language
English
Document Type
Product Information Sheet 2
Product Name
RosetteSep™ DM-L Density Medium
Catalog #
15705
Lot #
Lots above 1000053496
Language
Multi
Document Type
Safety Data Sheet
Product Name
RosetteSep™ DM-L Density Medium
Catalog #
15705
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All
Language
English
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 B Cell Research
Workflow Stages for Total Lymphocyte Research
Workflow Stages for T Cell Research
Workflow Stages for HLA Analysis
Resources and Publications
Educational Materials (3)
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.
Publications (1)
Completely phased genome sequencing through chromosome sorting. Yang H et al. Proceedings of the National Academy of Sciences of the United States of America 2011 JAN
Abstract
The two haploid genome sequences that a person inherits from the two parents represent the most fundamentally useful type of genetic information for the study of heritable diseases and the development of personalized medicine. Because of the difficulty in obtaining long-range phase information, current sequencing methods are unable to provide this information. Here, we introduce and show feasibility of a scalable approach capable of generating genomic sequences completely phased across the entire chromosome.
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