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EasySep™人记忆B细胞分选试剂盒

采用可解离磁珠对人记忆B细胞进行免疫磁珠分选

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产品号 #17864

人记忆B细胞的免疫磁珠正选

产品优势

  • 只需33分钟即可分离高纯度的人记忆B细胞
  • 无需清洗即可去除EasySep™ Releasable RapidSpheres™可解离磁珠
  • 可选择性地从同一样本中分离无标记的naïve B细胞

产品组分包括

  • EasySep™人记忆B细胞分选试剂盒（产品号 #17864）
    • EasySep™人记忆B细胞预富集抗体混合物，2 x 1 mL
    • EasySep™ Dextran RapidSpheres™ 磁珠，2 x 1 mL
    • EasySep™人CD27正选抗体混合物，1 mL
    • EasySep™ Releasable RapidSpheres™可解离磁珠，2 x 1 mL
    • EasySep™解离缓冲液，2 x 1 mL
main product photo
EasySep™人记忆B细胞分选试剂盒
概述
实验数据
产品说明书及文档
应用领域
相关材料与文献

概述

使用EasySep™人记忆B细胞分选试剂盒，可轻松从新鲜或冻存的人外周血单个核细胞（PBMCs）或白细胞单采术样本通过免疫磁珠正选分离出高纯度且无磁珠结合的人记忆B细胞。EasySep™结合了单克隆抗体的特异性和无柱磁性系统的简便性，迄今已广泛应用于发表的研究中超过20年。
该EasySep™正选通过抗体四聚体复合物和EasySep™可解离磁珠标记CD27+记忆B细胞。与常规的结合目的细胞的磁珠不同，这种磁珠具有可解离性。目的细胞先用抗体和磁珠标记，并通过EasySep™ 磁极进行无柱分选。非目的细胞通过简单倾倒弃去，而目的细胞则保留在分离管中。
随后，使用解离试剂将EasySep™分选出的CD27+细胞表面结合的磁珠进行去除。使用这款EasySep™解离分选试剂盒进行磁珠分选仅需40分钟，分选后的目的细胞即可立即用于下游实验。分选后目的细胞表面仍结合有抗体复合物，可能会与Billiant Violet™偶联抗体、聚乙二醇修饰
的蛋白或其他化学相关配体相互作用。
了解更多关于免疫磁珠EasySep™技术的工作原理，或直接选用经EasySep™人记忆B细胞分选试剂盒分离的、符合伦理规范的即用型冻存人外周血CD19+CD27+记忆B细胞。探索更多优化您实验流程的产品，包括培养基、添加剂、抗体等配套试剂。

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

实验数据

Typical EasySep™ Human Memory B Cell Isolation Profile

Figure 1. Typical EasySep™ Human Memory B Cell Isolation Profile

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).

Expansion and Viability of Human Memory B Cells Cultured with ImmunoCult™ Human B Cell Expansion Kit

Figure 2. Expansion and Viability of Human Memory B Cells Cultured with ImmunoCult™ Human B Cell Expansion Kit

Memory B cells were isolated from human PBMCs (leukopak) using EasySep™ Human Memory B Cell Isolation Kit and were seeded at 0.5 x 10⁵ cells/well in 48-well tissue culture plates and cultured with the ImmunoCult™ Human B Cell Expansion Kit. The cells were passaged every 2 - 4 days and the fold expansion of viable cells (A) and cell viability (B) were calculated at each timepoint. Data represent the mean ± 1 SD of triplicate cultures for the same donor.

产品说明书与文档

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

Document Type
Product Name
Catalog #
Lot #
Language
Document Type
Product Information Sheet
Product Name
EasySep™ Human Memory B Cell Isolation Kit
Catalog #
17864
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Product Name
EasySep™ Human Memory B Cell Isolation Kit
Catalog #
17864
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Product Name
EasySep™ Human Memory B Cell Isolation Kit
Catalog #
17864
Lot #
All
Language
English
Document Type
Safety Data Sheet 3
Product Name
EasySep™ Human Memory B Cell Isolation Kit
Catalog #
17864
Lot #
All
Language
English
Document Type
Safety Data Sheet 4
Product Name
EasySep™ Human Memory B Cell Isolation Kit
Catalog #
17864
Lot #
All
Language
English
Document Type
Safety Data Sheet 5
Product Name
EasySep™ Human Memory B Cell Isolation Kit
Catalog #
17864
Lot #
All
Language
English

应用领域

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

相关材料与文献

Educational Materials (11)

Human B Cell Isolation Product Selection
Brochure
Human B Cell Isolation Product Selection
EasySep™ Cell Separation Technology
Brochure
EasySep™ Cell Separation Technology
Tools for Your Immunology Research
Brochure
Tools for Your Immunology Research
Frequencies of Human Cell Types in Blood-Related Sources
Wallchart
Frequencies of Human Cell Types in Blood-Related Sources
Antigen Processing and Presentation
Wallchart
Antigen Processing and Presentation
Human Immune Cytokines
Wallchart
Human Immune Cytokines
How to Isolate PBMCs from Whole Blood Using Density Gradient Centrifugation (Ficoll™ or Lymphoprep™)
1:37
Video
How to Isolate PBMCs from Whole Blood Using Density Gradient Centrifugation (Ficoll™ or Lymphoprep...
Isolate Cells with a Simple Pour-Off: EasySep™ Cell Separation Technology
1:13
Video
Isolate Cells with a Simple Pour-Off: EasySep™ Cell Separation Technology
How EasySep™ Magnetic Cell Separation Technology Works: Fast and Easy Cell Isolation
1:57
Video
How EasySep™ Magnetic Cell Separation Technology Works: Fast and Easy Cell Isolation
Online Immunology Journal Club: Advances in Live-Attenuated Vaccine Development for Zika Virus
28:44
Webinar
Online Immunology Journal Club: Advances in Live-Attenuated Vaccine Development for Zika Virus
Fast and Easy Isolation of CD27-Positive Human Memory B Cells Using EasySep™ Releasable RapidSpheres™
Scientific Poster
Fast and Easy Isolation of CD27-Positive Human Memory B Cells Using EasySep™ Releasable RapidSpher...

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.
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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.
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