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Easily isolate highly purified and magnetic particle-free human cells labeled with biotinylated antibodies from fresh or previously frozen human peripheral blood mononuclear cells (PBMCs) and washed leukapheresis samples, by immunomagnetic positive selection, with the EasySep? Release Human Biotin Positive Selection Kit. Widely used in published research for more than 20 years, EasySep? combines the specificity of monoclonal antibodies with the simplicity of a column-free magnetic system.
In this EasySep? positive selection procedure, desired cells are first labeled with antibody complexes recognizing biotin and magnetic particles called EasySep? Releasable RapidSpheres?. Unlike traditional magnetic particles, which stay bound to the target cells, these RapidSpheres? have a releasable feature. After separation using an EasySep? magnet, bound magnetic particles are removed from the EasySep?-isolated, biotin antibody-labeled cells using a release agent, and unwanted cells are targeted for depletion. The final isolated fraction contains highly purified biotin+ cells that are immediately ready for downstream applications such as flow cytometry, culture, or DNA/RNA extraction. Following cell isolation with this EasySep? kit, antibody complexes remain bound to the cell surface and may interact with Brilliant Violet? antibody conjugates, polyethylene glycol-modified proteins, or other chemically related ligands.
This product replaces the EasySep? Human Biotin Positive Selection Kit (Catalog #18553), providing highly purified particle-free cells.
Learn more about how immunomagnetic EasySep? technology works. Explore additional products optimized for your workflow, including those for culture media, supplements, antibodies, and more.
Magnet Compatibility
? EasySep? Magnet (Catalog #18000)
? “The Big Easy” EasySep? Magnet (Catalog #18001)
? EasyPlate? Magnet (Catalog #18102)
? EasyEights? Magnet (Catalog #18103)
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
Figure 1. Typical EasySep? Release Human Biotin Positive Selection Profile with PBMCs
Starting with fresh PBMCs, the purities of the start and final isolated fractions are 34.6% and 97.1%, respectively, using a biotinylated anti-human CD45RO
antibody and EasySep? Release Human Biotin Positive Selection Kit (as assessed by labeling with CD45RO and CD45RA).
Figure 2. Typical EasySep? Release Human Biotin Positive Selection Profile with Mouse Splenocytes
Starting with mouse splenocytes, the purities of the start and final isolated fractions are 58.4% and 95.1%, respectively, using a biotinylated anti-mouse CD19 antibody and EasySep? Release Mouse Biotin Positive Selection Kit (as assessed by labeling with CD19 and CD45R/B220).
Regulatory Programs of B-cell Activation and Germinal Center Reaction Allow B-ALL Escape from CD19 CAR T-cell Therapy.
N. G. Im et al.
Cancer immunology research 2022 sep
Abstract
Chimeric antigen receptor (CAR) T-cell therapy has led to tremendous successes in the treatment of B-cell malignancies. However, a large fraction of treated patients relapse, often with disease expressing reduced levels of the target antigen. Here, we report that exposing CD19+ B-cell acute lymphoblastic leukemia (B-ALL) cells to CD19 CAR T cells reduced CD19 expression within hours. Initially, CD19 CAR T cells caused clustering of CD19 at the T cell-leukemia cell interface followed by CD19 internalization and decreased CD19 surface expression on the B-ALL cells. CD19 expression was then repressed by transcriptional rewiring. Using single-cell RNA sequencing and single-cell assay for transposase-accessible chromatin using sequencing, we demonstrated that a subset of refractory CD19low cells sustained decreased CD19 expression through transcriptional programs of physiologic B-cell activation and germinal center reaction. Inhibiting B-cell activation programs with the Bruton's tyrosine kinase inhibitor ibrutinib increased the cytotoxicity of CD19 CAR T cells without affecting CAR T-cell viability. These results demonstrate transcriptional plasticity as an underlying mechanism of escape from CAR T cells and highlight the importance of combining CAR T-cell therapy with targeted therapies that aim to overcome this plasticity. See related Spotlight by Zhao and Melenhorst, p. 1040.
EasySep? Release Human Biotin Positive Selection Kit
New look, same high quality and support! You may notice that your instrument or reagent packaging looks slightly different from images displayed on the website, or from previous orders. We are updating our look but rest assured, the products themselves and how you should use them have not changed. Learn more
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