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 -
Label for EasySep™ Human Memory B Cell Isolation Kit
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Required Products
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Magnet for column-free immunomagnetic separation -
Cell separation buffer -
Compatible antibodies for purity assessment of isolated cells
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.
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
• No-wash removal of EasySep™ Releasable RapidSpheres™
• Optional isolation of untouched naïve B cells from the same sample
Components:
- 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
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
Related Products
Labeling Antibodies
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Anti-Human CD19 Antibody, Clone HIB19
Mouse monoclonal IgG1 antibody against human, chimpanzee CD19 -
Anti-Mouse CD27 Antibody, Clone LG.3A10
Hamster (Armenian) monoclonal IgG1 antibody against human, mouse, rat CD27
Scientific Resources
Product Documentation
Document Type
Product Name
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Educational Materials
(9)
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
Data
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).
Publications
(1)
Cell 2020
Potent Neutralizing Antibodies against SARS-CoV-2 Identified by High-Throughput Single-Cell Sequencing of Convalescent Patients' B Cells.
Abstract
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.
PRODUCTS ARE FOR RESEARCH USE ONLY AND NOT INTENDED FOR HUMAN OR ANIMAL DIAGNOSTIC OR THERAPEUTIC USES UNLESS OTHERWISE STATED. FOR ADDITIONAL INFORMATION ON QUALITY AT STEMCELL, REFER TO WWW.STEMCELL.COM/COMPLIANCE.