EasySep™ Human Memory B Cell Isolation Kit

Immunomagnetic cell isolation kit using particle release technology

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EasySep™ Human Memory B Cell Isolation Kit

Immunomagnetic cell isolation kit using particle release technology

From: 1,121 USD
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Immunomagnetic cell isolation kit using particle release technology
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Product Advantages


  • Highly purified human memory B cells isolated in as little as 33 minutes

  • No-wash removal of EasySep™ Releasable RapidSpheres™

  • Optional isolation of untouched naïve B cells from the same sample

What's Included

  • EasySep™ Human Memory B Cell Isolation Kit (Catalog #17864)
    • EasySep™ Human Memory B Cell Pre-Enrichment Cocktail, 2 x 1 mL
    • EasySep™ Dextran RapidSpheres™, 2 x 1 mL
    • EasySep™ Human CD27 Positive Selection Cocktail, 1 mL
    • EasySep™ Releasable RapidSpheres™, 2 x 1 mL
    • EasySep™ Release Buffer, 2 x 1 mL

Overview

The EasySep™ Human Memory B Cell Isolation Kit is a two-step method designed to isolate CD27+ B cells from fresh or previously frozen peripheral blood mononuclear cells. First, CD27+ cells are isolated by column-free immunomagnetic positive selection using antibody complexes and EasySep™ Releasable RapidSpheres™. Then, bound magnetic particles are removed from the EasySep™-isolated CD27+ cells, and unwanted non-B cells are targeted for depletion using antibody complexes and EasySep™ Dextran RapidSpheres™. The final isolated fraction contains highly purified CD27+ memory B cells that are immediately ready for downstream applications. An optional protocol allows for the isolation of naïve B cells in parallel for use in functional studies.
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
PBMC
Selection Method
Positive
Application
Cell Isolation
Brand
EasySep
Area of Interest
Immunology

Data Figures

Starting with PBMCs, the memory B cell content (CD19+CD27+) of the isolated fraction is typically 97 ± 2% (mean ± SD using the purple EasySep™ Magnet). Using the optional protocol, the naïve B cell content (CD19+CD27-) of the isolated fraction is typically 93 ± 5% (mean ± SD).

Protocols and Documentation

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

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17864
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English
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Safety Data Sheet 1
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Safety Data Sheet 2
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17864
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Safety Data Sheet 3
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Safety Data Sheet 4
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Safety Data Sheet 5
Catalog #
17864
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English

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)

Potent Neutralizing Antibodies against SARS-CoV-2 Identified by High-Throughput Single-Cell Sequencing of Convalescent Patients' B Cells. Y. Cao et al. Cell 2020

Abstract

The COVID-19 pandemic urgently needs therapeutic and prophylactic interventions. Here, we report the rapid identification of SARS-CoV-2-neutralizing antibodies by high-throughput single-cell RNA and VDJ sequencing of antigen-enriched B cells from 60 convalescent patients. From 8,558 antigen-binding IgG1+ clonotypes, 14 potent neutralizing antibodies were identified, with the most potent one, BD-368-2, exhibiting an IC50 of 1.2 and 15 ng/mL against pseudotyped and authentic SARS-CoV-2, respectively. BD-368-2 also displayed strong therapeutic and prophylactic efficacy in SARS-CoV-2-infected hACE2-transgenic mice. Additionally, the 3.8 {\AA} cryo-EM structure of a neutralizing antibody in complex with the spike-ectodomain trimer revealed the antibody's epitope overlaps with the ACE2 binding site. Moreover, we demonstrated that SARS-CoV-2-neutralizing antibodies could be directly selected based on similarities of their predicted CDR3H structures to those of SARS-CoV-neutralizing antibodies. Altogether, we showed that human neutralizing antibodies could be efficiently discovered by high-throughput single B cell sequencing in response to pandemic infectious diseases.
FcγRIIB-Independent Mechanisms Controlling Membrane Localization of the Inhibitory Phosphatase SHIP in Human B Cells. Pauls SD et al. Journal of immunology (Baltimore, Md. : 1950) 2016 JUL

Abstract

SHIP is an important regulator of immune cell signaling that functions to dephosphorylate the phosphoinositide phosphatidylinositol 3,4,5-trisphosphate at the plasma membrane and mediate protein-protein interactions. One established paradigm for SHIP activation involves its recruitment to the phospho-ITIM motif of the inhibitory receptor FcγRIIB. Although SHIP is essential for the inhibitory function of FcγRIIB, it also has critical modulating functions in signaling initiated from activating immunoreceptors such as B cell Ag receptor. In this study, we found that SHIP is indistinguishably recruited to the plasma membrane after BCR stimulation with or without FcγRIIB coligation in human cell lines and primary cells. Interestingly, fluorescence recovery after photobleaching analysis reveals differential mobility of SHIP-enhanced GFP depending on the mode of stimulation, suggesting that although BCR and FcγRIIB can both recruit SHIP, this occurs via distinct molecular complexes. Mutagenesis of a SHIP-enhanced GFP fusion protein reveals that the SHIP-Src homology 2 domain is essential in both cases whereas the C terminus is required for recruitment via BCR stimulation, but is less important with FcγRIIB coligation. Experiments with pharmacological inhibitors reveal that Syk activity is required for optimal stimulation-induced membrane localization of SHIP, whereas neither PI3K or Src kinase activity is essential. BCR-induced association of SHIP with binding partner Shc1 is dependent on Syk, as is tyrosine phosphorylation of both partners. Our results indicate that FcγRIIB is not uniquely able to promote membrane recruitment of SHIP, but rather modulates its function via formation of distinct signaling complexes. Membrane recruitment of SHIP via Syk-dependent mechanisms may be an important factor modulating immunoreceptor signaling.
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