EasySep™ Direct Human Naïve B Cell Isolation Kit

Immunomagnetic negative selection kit from whole blood
概要
This kit is used to isolate highly purified naïve B cells directly from human whole blood. Isolated cells are unlabeled with antibodies or particles and undergo minimal manipulation.

This kit targets non-naïve B cells for removal with antibodies recognizing specific surface markers. Unwanted cells are labeled with antibodies and EasySep™ Direct RapidSpheres™, and separated using an EasySep™ magnet. Desired cells are simply collected into a new tube and are immediately available for downstream applications such as flow cytometry, culture or DNA/RNA extraction.
Advantages
• > 99.9% red blood cell (RBC) depletion without the need for density gradient centrifugation, sedimentation, or lysis
• Fast, easy-to-use and column-free
• Up to 95% purity of isolated cells
• Isolated cells are untouched
Components
  • EasySep™ Direct Human Naïve B Cell Isolation Kit (Catalog #19264)
    • EasySep™ Direct Human Naïve B Cell Isolation Cocktail, 2 x 2.5 mL
    • EasySep™ Direct RapidSpheres™, 4 x 2.5 mL
Magnet Compatibility
• EasySep™ Magnet (Catalog #18000)
• “The Big Easy” EasySep™ Magnet (Catalog #18001)
• Easy 50 EasySep™ Magnet (Catalog #18002)
• EasyEights™ EasySep™ Magnet (Catalog #18103)
Subtype
Cell Isolation Kits
Cell Type
B Cells
Species
Human
Sample Source
Whole Blood
Selection Method
Negative
Application
Cell Isolation
Brand
EasySep
Area of Interest
Immunology
数据及文献

Data

Starting with human whole blood from normal healthy donors, the typical naïve B cell (CD19+CD27-) content of the non-lysed final isolated fraction is 918 ± 36% (gated on CD45) or 824 ± 126% (not gated on CD45)

Figure 1. Typical EasySep™ Direct Human Naïve B Cell Isolation Profile

Starting with human whole blood from normal healthy donors, the typical naïve B cell (CD19+CD27-) content of the non-lysed final isolated fraction is 91.8 ± 3.6% (gated on CD45) or 82.4 ± 12.6% (not gated on CD45). In the above example, the naïve B cell (CD19+CD27-) content of the lysed whole blood start sample and non-lysed final isolated fraction is 5.1% and 91.5% (gated on CD45), respectively, or 5.1% and 90.0% (not gated on CD45), respectively. The starting frequency of naïve B cells in the non-lysed whole blood start sample above is 0.009% (data not shown).

Publications (1)

Scientific reports 2019 jan BCR-associated factors driving chronic lymphocytic leukemia cells proliferation ex vivo. C. Schleiss et al.
Abstract
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Abstract

A chronic antigenic stimulation is believed to sustain the leukemogenic development of chronic lymphocytic leukemia (CLL) and most of lymphoproliferative malignancies developed from mature B cells. Reproducing a proliferative stimulation ex vivo is critical to decipher the mechanisms of leukemogenesis in these malignancies. However, functional studies of CLL cells remains limited since current ex vivo B cell receptor (BCR) stimulation protocols are not sufficient to induce the proliferation of these cells, pointing out the need of mandatory BCR co-factors in this process. Here, we investigated benefits of several BCR co-stimulatory molecules (IL-2, IL-4, IL-15, IL-21 and CD40 ligand) in multiple culture conditions. Our results demonstrated that BCR engagement (anti-IgM ligation) concomitant to CD40 ligand, IL-4 and IL-21 stimulation allowed CLL cells proliferation ex vivo. In addition, we established a proliferative advantage for ZAP70 positive CLL cells, associated to an increased phosphorylation of ZAP70/SYK and STAT6. Moreover, the use of a tri-dimensional matrix of methylcellulose and the addition of TLR9 agonists further increased this proliferative response. This ex vivo model of BCR stimulation with T-derived cytokines is a relevant and efficient model for functional studies of CLL as well as lymphoproliferative malignancies.
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