RosetteSep™ DM-L Density Medium

Density gradient medium

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RosetteSep™ DM-L Density Medium

Density gradient medium for use with RosetteSep™ CD3+ T cell, CD4+ T cell, CD8+ T cell, and B cell enrichment cocktails

100 mL
Catalog #15705
61 USD

Overview

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. 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.
Contains:
• Iodixanol
• Hetastarch
• Sodium chloride
• Sodium lactate
• Dextrose
• Calcium chloride
• Potassium chloride
• Magnesium chloride
• Water
Subtype:
Density Gradient Media
Cell Type:
B Cells; T Cells
Species:
Human
Sample Source:
Buffy Coat; Whole Blood
Selection Method:
Negative
Application:
In Vitro Diagnostic; Cell Isolation
Brand:
RosetteSep
Area of Interest:
Immunology

Scientific Resources

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.
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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. Minimal Granulocyte Contamination Using RosetteSep™ DM-L Density Medium

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)

Rosettesep™ DM-L Improves the Recovery of Lymphocytes Enriched with Rosettesep™ from Fresh Blood

Figure 2. Rosettesep™ DM-L Improves the Recovery of Lymphocytes Enriched with Rosettesep™ from Fresh Blood

Cell Purity Is Equivalent or Higher for Rosettesep™ Lymphocyte Enrichment Using Rosettesep™ DM-L Instead of Ficoll-Paque

Figure 3. Cell Purity Is Equivalent or Higher for Rosettesep™ Lymphocyte Enrichment Using Rosettesep™ DM-L Instead of Ficoll-paque®

Publications

(1)
Proceedings of the National Academy of Sciences of the United States of America 2011 JAN

Completely phased genome sequencing through chromosome sorting.

Yang H et al.

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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