# RosetteSep™ Human CD4 Depletion Cocktail

Immunodensity depletion cocktail

From: 101 USD

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Immunodensity depletion cocktail
From: 101 USD

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# Overview

The RosetteSep™ Human CD4 Depletion Cocktail is designed to deplete CD4+ cells from whole blood. Unwanted cells are targeted for removal with Tetrameric Antibody Complexes recognizing CD4 and glycophorin A on red blood cells (RBCs). When centrifuged over a buoyant density medium such as Lymphoprep™ (Catalog #07801), the unwanted cells pellet along with the RBCs. The CD4+ depleted fraction is present as a highly enriched population at the interface between the plasma and the buoyant density medium.
• 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 CD4 Depletion Cocktail (Catalog #15622)
• RosetteSep™ Human CD4 Depletion Cocktail, 2 mL
• RosetteSep™ Human CD4 Depletion Cocktail (Catalog #15662)
• RosetteSep™ Human CD4 Depletion Cocktail, 5 x 2 mL
Subtype:
Cell Isolation Kits
Cell Type:
T Cells; T Cells, CD4+
Species:
Human
Sample Source:
Buffy Coat; Whole Blood
Selection Method:
Depletion
Application:
Cell Isolation
Brand:
RosetteSep
Area of Interest:
Immunology

Document Type
Product Name
Catalog #
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(4)

# 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.

# 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.

Research Area Workflow Stages for
Workflow Stages

# Data and Publications

## Publications

(2)
Journal of immunology (Baltimore, Md. : 1950) 2019 jul

### Metformin Inhibits the Type 1 IFN Response in Human CD4+ T Cells.

A. A. Titov et al.
Abstract

### Abstract

In systemic lupus erythematosus, defective clearance of apoptotic debris and activation of innate cells result in a chronically activated type 1 IFN response, which can be measured in PBMCs of most patients. Metformin, a widely used prescription drug for Type 2 diabetes, has a therapeutic effect in several mouse models of lupus through mechanisms involving inhibition of oxidative phosphorylation and a decrease in CD4+ T cell activation. In this study, we report that in CD4+ T cells from human healthy controls and human systemic lupus erythematosus patients, metformin inhibits the transcription of IFN-stimulated genes (ISGs) after IFN-alpha treatment. Accordingly, metformin inhibited the phosphorylation of pSTAT1 (Y701) and its binding to IFN-stimulated response elements that control ISG expression. These effects were independent of AMPK activation or mTORC1 inhibition but were replicated using inhibitors of the electron transport chain respiratory complexes I, III, and IV. This indicates that mitochondrial respiration is required for ISG expression in CD4+ T cells and provides a novel mechanism by which metformin may exert a therapeutic effect in autoimmune diseases.
Mechanisms of development 2019

### Developmental regulation of Wnt signaling by Nagk and the UDP-GlcNAc salvage pathway.

L. R. Neitzel et al.
Abstract

### Abstract

In a screen for human kinases that regulate Xenopus laevis embryogenesis, we identified Nagk and other components of the UDP-GlcNAc glycosylation salvage pathway as regulators of anteroposterior patterning and Wnt signaling. We find that the salvage pathway does not affect other major embryonic signaling pathways (Fgf, TGF$\beta$, Notch, or Shh), thereby demonstrating specificity for Wnt signaling. We show that the role of the salvage pathway in Wnt signaling is evolutionarily conserved in zebrafish and Drosophila. Finally, we show that GlcNAc is essential for the growth of intestinal enteroids, which are highly dependent on Wnt signaling for growth and maintenance. We propose that the Wnt pathway is sensitive to alterations in the glycosylation state of a cell and acts as a nutritional sensor in order to couple growth/proliferation with its metabolic status. We also propose that the clinical manifestations observed in congenital disorders of glycosylation (CDG) in humans may be due, in part, to their effects on Wnt signaling during development.
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