EasySep™ Human Plasmacytoid DC Isolation Kit

Immunomagnetic negative selection kit

EasySep™ Human Plasmacytoid DC Isolation Kit

Immunomagnetic negative selection kit

From: 1,134 USD
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Immunomagnetic negative selection kit
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Product Advantages


  • Fast, easy-to-use, and column-free

  • Up to 97% purity

  • Untouched, viable cells

What's Included

  • EasySep™ Human Plasmacytoid DC Isolation Kit (Catalog #17977)
    • EasySep™ Human Plasmacytoid DC Isolation Cocktail Component A, 2 x 1 mL
    • EasySep™ Human Plasmacytoid DC Isolation Cocktail Component B, 2 x 1 mL
    • EasySep™ Dextran RapidSpheres™ 50103, 4 x 1 mL
    • Anti-Human CD32 (Fc gamma RII) Blocker, 2 x 0.8 mL

Overview

The EasySep™ Human Plasmacytoid DC Isolation Kit is designed to isolate plasmacytoid dendritic cells (pDCs) from human peripheral blood mononuclear cells (PBMCs) or leukapheresis by immunomagnetic negative selection. The EasySep™ procedure involves labeling unwanted cells with antibody complexes and magnetic particles. The magnetically labeled cells are separated from the untouched desired cells by using an EasySep™ magnet and simply pouring or pipetting the desired cells into a new tube.

This product replaces the EasySep™ Human Plasmacytoid DC Enrichment Kit (Catalog #19062), for even faster cell isolations and is compatible with a greater number of EasySep™ magnets.
Magnet Compatibility
• EasySep™ Magnet (Catalog #18000)
• “The Big Easy” EasySep™ Magnet (Catalog #18001)
• EasyEights™ EasySep™ Magnet (Catalog #18103)
• Easy 50 EasySep™ Magnet (Catalog #18002)
• RoboSep™-S (Catalog #21000)
Subtype
Cell Isolation Kits
Cell Type
Dendritic Cells
Species
Human
Sample Source
Leukapheresis, PBMC
Selection Method
Negative
Application
Cell Isolation
Brand
EasySep, RoboSep
Area of Interest
Immunology, Infectious Diseases

Data Figures

FACS Profile Results with EasySep™ Human Plasmacytoid DC Isolation Kit

Figure 1. Typical EasySep™ Human Plasmacytoid DC Isolation Profile

Starting with leukapheresis samples, the pDC content (Lin-HLA-DR+CD123+) of the isolated fraction is typically 90 ± 5.3% (mean ± SD using the silver “Big Easy” EasySep™ Magnet). In the above example, the purities of the start and final isolated fractions are 0.2% and 89.8%, respectively.

Protocols and Documentation

Find supporting information and directions for use in the Product Information Sheet or explore additional protocols below.

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Safety Data Sheet 1
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Safety Data Sheet 5
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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.

Resources and Publications

Publications (2)

HMGB1 Is Involved in IFN-α Production and TRAIL Expression by HIV-1-Exposed Plasmacytoid Dendritic Cells: Impact of the Crosstalk with NK Cells. Sa&iuml et al. PLoS pathogens 2016 FEB

Abstract

Plasmacytoid dendritic cells (pDCs) are innate sensors of viral infections and important mediators of antiviral innate immunity through their ability to produce large amounts of IFN-α. Moreover, Toll-like receptor 7 (TLR7) and 9 (TLR9) ligands, such as HIV and CpG respectively, turn pDCs into TRAIL-expressing killer pDCs able to lyse HIV-infected CD4+ T cells. NK cells can regulate antiviral immunity by modulating pDC functions, and pDC production of IFN-α as well as cell-cell contact is required to promote NK cell functions. Impaired pDC-NK cell crosstalk was reported in the setting of HIV-1 infection, but the impact of HIV-1 on TRAIL expression and innate antiviral immunity during this crosstalk is unknown. Here, we report that low concentrations of CCR5-tropic HIV-1Ba-L promote the release of pro-inflammatory cytokines such as IFN-α, TNF-α, IFN-γ and IL-12, and CCR5-interacting chemokines (MIP-1α and MIP-1β) in NK-pDCs co-cultures. At high HIV-1BaL concentrations, the addition of NK cells did not promote the release of these mediators, suggesting that once efficiently triggered by the virus, pDCs could not integrate new activating signals delivered by NK cells. However, high HIV-1BaL concentrations were required to trigger IFN-α-mediated TRAIL expression at the surface of both pDCs and NK cells during their crosstalk. Interestingly, we identified the alarmin HMGB1, released at pDC-NK cell synapse, as an essential trigger for the secretion of IFN-α and IFN-related soluble mediators during the interplay of HIV-1 exposed pDCs with NK cells. Moreover, HMGB1 was found crucial for mTRAIL translocation to the plasma membrane of both pDCs and NK cells during their crosstalk following pDC exposure to HIV-1. Data from serum analyses of circulating HMGB1, HMGB1-specific antibodies, sTRAIL and IP-10 in a cohort of 67 HIV-1+ patients argue for the in vivo relevance of these observations. Altogether, these findings identify HMGB1 as a trigger for IFN-α-mediated TRAIL expression at the surface of pDCs and NK cells, and they suggest a novel mechanism of innate control of HIV-1 infection.
Plasmacytoid dendritic cells reduce HIV production in elite controllers. Machmach K et al. Journal of virology 2012 APR

Abstract

HIV elite controllers (EC) are a rare group of HIV-infected patients who are able to maintain undetectable viral loads during a long period of time in the absence of antiretroviral treatment. Adaptive immunity and host genetic factors, although implicated, do not entirely explain this phenomenon. On the other hand, plasmacytoid dendritic cells (pDCs) are the principal type I interferon (IFN) producers in response to viral infection, and it is unknown whether pDCs are involved in the control of HIV infection in EC. In our study, we analyzed peripheral pDC levels and IFN-α production by peripheral blood mononuclear cells (PBMCs) in EC compared to other groups of HIV-infected patients, the ability of pDCs to reduce HIV production in vitro, and the mechanisms potentially involved. We showed preserved pDC counts and IFN-α production in EC. We also observed a higher capacity of pDCs from EC to reduce HIV production and to induce T cell apoptosis, whereas pDCs from viremic patients barely responded without previous Toll-like receptor 9 (TLR-9) stimulus. The preserved functionality of pDCs from EC to reduce viral production may be one of the mechanisms involved in the control of HIV viremia in these subjects. These results demonstrate the importance of innate immunity in HIV pathogenesis, and an understanding of pDC mechanisms would be helpful for the design of new therapies.