RosetteSep™ Human Custom Cocktail

Immunodensity negative selection cocktail
RosetteSep™ Human Custom Cocktail

Immunodensity isolation of untouched human cells

As requested
Catalog # 15309
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Required Products
  1. Lymphoprep™|07851
    Lymphoprep™

    Density gradient medium for the isolation of mononuclear cells

Overview

The RosetteSep™ Human Custom Cocktail is designed to deplete unwanted cells from fresh whole blood by negative selection. When centrifuged over a buoyant density medium such as Lymphoprep™ (Catalog #07801), the unwanted cells pellet along with the RBCs. The cells of interest are present as an enriched population at the interface between the plasma and the buoyant density medium. STEMCELL Technologies offers a wide selection of antibodies for custom separation and will work with you to isolate a cell type of interest. Please contact our technical support team at techsupport@stemcell.com for more information.
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
Components
  • RosetteSep™ Human Custom Cocktail (Catalog #15309)
    • RosetteSep™ Human Custom Enrichment Cocktail
Magnet Compatibility
 
Subtype
Cell Isolation Kits
Cell Type
B Cells, Dendritic Cells, Granulocytes and Subsets, Hematopoietic Stem and Progenitor Cells, Macrophages, Marrow Stromal Cells, Mesenchymal Stem and Progenitor Cells, Monocytes, Mononuclear Cells, Myeloid Cells, NK Cells, Other, Plasma, T Cells
Species
Human
Sample Source
Buffy Coat, Whole Blood
Selection Method
Negative
Application
Cell Isolation
Brand
RosetteSep
Area of Interest
Immunology, Stem Cell Biology

Scientific Resources

Product Documentation

Document Type Product Name Catalog # Lot # Language
Document Type
Product Information Sheet
Product Name
RosetteSep™ Human Custom Cocktail
Catalog #
15309
Lot #
All
Language
English

Educational Materials (4)

Brochure
Tools For Your Immunology Research
Wallchart
Human Immune Cytokines
Video
Isolate Cells from Whole Blood without Columns or Magnets: RosetteSep™ Immunodensity Cell Separation
2:19
Isolate Cells from Whole Blood without Columns or Magnets: RosetteSep™ Immunodensity Cell Separation
Video
Isolate Highly Purified Cells for HLA Analysis with RosetteSep™ Immunodensity Cell Isolation
1:21
Isolate Highly Purified Cells for HLA Analysis with RosetteSep™ Immunodensity Cell Isolation

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

Data and Publications

Publications (1)

Cell death {\&} disease 2019 Combination of 5-fluorouracil and thymoquinone targets stem cell gene signature in colorectal cancer cells. B. Ndreshkjana et al.

Abstract

Cancer stem cells (CSCs) residing in colorectal cancer tissues have tumorigenic capacity and contribute to chemotherapeutic resistance and disease relapse. It is well known that the survival of colorectal CSCs after 5-fluorouracil (5-FU)-based therapy leads to cancer recurrence. Thus CSCs represent a promising drug target. Here, we designed and synthesized novel hybrid molecules linking 5-FU with the plant-derived compound thymoquinone (TQ) and tested the potential of individual compounds and their combination to eliminate colorectal CSCs. Both, Combi and SARB hybrid showed augmented cytotoxicity against colorectal cancer cells, but were non-toxic to organoids prepared from healthy murine small intestine. NanoString analysis revealed a unique signature of deregulated gene expression in response to the combination of TQ and 5-FU (Combi) and SARB treatment. Importantly, two principle stem cell regulatory pathways WNT/{\ss}-Catenin and PI3K/AKT were found to be downregulated after Combi and hybrid treatment. Furthermore, both treatments strikingly eliminated CD133+ CSC population, accompanying the depleted self-renewal capacity by eradicating long-term propagated 3D tumor cell spheres at sub-toxic doses. In vivo xenografts on chicken eggs of SARB-treated HCT116 cells showed a prominent nuclear {\ss}-Catenin and E-cadherin staining. This was in line with the reduced transcriptional activity of {\ss}-Catenin and diminished cell adhesion under SARB exposure. In contrast to 5-FU, both, Combi and SARB treatment effectively reduced the angiogenic capacity of the remaining resistant tumor cells. Taken together, combination or hybridization of single compounds target simultaneously a broader spectrum of oncogenic pathways leading to an effective eradication of colorectal cancer cells.
View All Publications

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