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EasySep™人B细胞分选试剂盒

免疫磁珠负选人B细胞
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产品号 #（选择产品）

产品号 #17954_C

免疫磁珠负选人B细胞

产品优势

  • 操作简单、快捷，且无需分离柱
  • 纯度高达96 %，回收率高
  • 分选得到的细胞不带标记

产品组分包括

  • EasySep™人B 细胞分选试剂盒（产品号 #17954）
    • EasySep™ 人B细胞分选抗体混合物, 1 mL
    • EasySep™ Dextran RapidSpheres™磁珠, 1 mL
    • EasySep™分选抗体混合物增强剂, 1 mL    
  • EasySep™人B 细胞分选试剂盒（产品号 #100-0971）
    • EasySep™ 人B细胞抗体混合物 ，1 x 10 mL（产品号 #300-0510）
    • EasySep™ 分选抗体混合物增强剂，1 x 10 mL（产品号 #300-0511）
    • EasySep™Dextran RapidSpheres™，1 x 10 mL（产品号 #300-0380）
  • RoboSep™ 人B细胞分选试剂盒（产品号 #17954RF）
    • EasySep™ 人B细胞分选抗体混合物, 1 mL
    • EasySep™ Dextran RapidSpheres™磁珠, 1 mL
    • EasySep™分选抗体混合物增强剂, 1 mL    
    • RoboSep™ 缓冲液（产品号 #20104）
    • RoboSep™过滤吸头（产品号 #20125）
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  1. EasySep™磁极
    EasySep™磁极
  2. EasySep™缓冲液
    EasySep™缓冲液
总览
实验数据
产品说明书及文档
应用领域
相关材料与文献
相关产品

总览

使用 EasySep™ 人B细胞分离试剂盒，可轻松高效地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中通过免疫磁珠负选获得高纯度的人B细胞。EasySep™技术结合单克隆抗体的特异性和免磁柱系统的简便性，已在发表的研究中广泛应用超过20年。

在该  EasySep™负 选流程中，非目的细胞会被抗体复合物与磁珠标记。表达以下标记物的非目的细胞将被靶向去除：CD2、CD3、CD14、CD16、CD36、CD43、CD56、CD66b和 GlyA。通过EasySep™磁极将被磁珠标记的细胞与未被标记的目的细胞分离，只需将目的细胞倾倒或移液至新试管中。磁珠分选最快仅需9分钟，所得目的B细胞即可用于流式细胞术、培养或DNA/RNA提取等下游应用。

该试剂盒可替代EasySep™人B细胞富集试剂盒 (产品号 #19054) 以实现更快速地细胞分选。

如需从白细胞单采术样本中大规模分选人B细胞，请选用大包装（1x1010 细胞）试剂盒（产品号  #100-0971）。

了解更多关于免疫磁珠EasySep™技术的工作原理，或如何通过RoboSep™实现免疫磁珠细胞分选全自动化。或直接选用 EasySep™ 人B细胞分选试剂盒提供的即用型、符合伦理道德的原代人外周血B细胞。探索其他针对您的工作流程优化的产品，包括培养基、补充剂、抗体等。     

 

磁极兼容性
• EasySep™磁极（产品号 #18000）
• “The Big Easy” EasySep™磁极（产品号 #18001）
• Easy 50 EasySep™磁极（产品号 #18002）
• EasyEights™ EasySep™磁极（产品号 #18103）
• RoboSep™-S（产品号 #21000）
• Easy 250 EasySep™磁极（产品号 #100-0821）
 
分类
细胞分选试剂盒
 
细胞类型
B 细胞
 
种属
人
 
样本来源
白细胞单采术样本、PBMC
 
分选方法
负选
 
应用
细胞分选
 
品牌
EasySep，RoboSep
 
研究领域
嵌合体，HLA，免疫
 

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实验数据

Figure 1. Typical EasySep™ Human B Cell Isolation Profile

Starting with human PBMCs, the B cell (CD3-CD19+) content of the isolated fraction is typically 95.1 ± 1.4% (mean ± SD). In the example above, the final purities of the start and isolated fractions are 4.5% and 94.9%, respectively.

产品说明书及文档

请在《产品说明书》中查找相关支持信息和使用说明，或浏览下方更多实验方案。

Document Type
Product Name
Catalog #
Lot #
Language
Document Type
产品说明书
Product Name
RoboSep™ 人B细胞分选试剂盒
Catalog #
17954RF
Lot #
All
Language
中文
Document Type
产品说明书
Product Name
EasySep™人B细胞分选试剂盒
Catalog #
17954
Lot #
All
Language
中文
Document Type
Product Information Sheet - with Easy 250 EasySep™ Magnet
Product Name
EasySep™ Human B Cell Isolation Kit
Catalog #
100-0971
Lot #
All
Language
English
Document Type
Product Information Sheet
Product Name
EasySep™ Human B Cell Isolation Kit
Catalog #
17954
Lot #
All
Language
English
Document Type
Product Information Sheet
Product Name
RoboSep™ Human B Cell Isolation Kit
Catalog #
17954RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Product Name
EasySep™ Human B Cell Isolation Kit
Catalog #
100-0971
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Product Name
EasySep™ Human B Cell Isolation Kit
Catalog #
100-0971
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Product Name
EasySep™ Human B Cell Isolation Kit
Catalog #
17954
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Product Name
EasySep™ Human B Cell Isolation Kit
Catalog #
17954
Lot #
All
Language
English
Document Type
Safety Data Sheet 3
Product Name
EasySep™ Human B Cell Isolation Kit
Catalog #
17954
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Product Name
RoboSep™ Human B Cell Isolation Kit
Catalog #
17954RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Product Name
RoboSep™ Human B Cell Isolation Kit
Catalog #
17954RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 3
Product Name
RoboSep™ Human B Cell Isolation Kit
Catalog #
17954RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 4
Product Name
RoboSep™ Human B Cell Isolation Kit
Catalog #
17954RF
Lot #
All
Language
English

应用领域

本产品专为以下研究领域设计，适用于工作流程中的高亮阶段。探索这些工作流程，了解更多我们为各研究领域提供的其他配套产品。

Research Area
Workflow Stages

相关材料与文献

技术资料 (13)

血液相关来源中人细胞类型的比例
挂图
血液相关来源中人细胞类型的比例
EasySep™细胞分选技术 - 更高效地分离细胞
产品手册
EasySep™细胞分选技术 - 更高效地分离细胞
Human B Cell Isolation Product Selection
产品手册
Human B Cell Isolation Product Selection
Isolate Human Immune Cells
产品手册
Isolate Human Immune Cells
细胞分选相关工具
产品手册
细胞分选相关工具
How to Isolate PBMCs from Whole Blood Using Density Gradient Centrifugation (Ficoll™ or Lymphoprep™)
视频
How to Isolate PBMCs from Whole Blood Using Density Gradient Centrifugation (Ficoll™ or Lymphoprep™)
Simultaneous Cell Isolation from Multiple Samples Using the EasyEights™ EasySep™ Magnet
视频
Simultaneous Cell Isolation from Multiple Samples Using the EasyEights™ EasySep™ Magnet
How to Isolate Cells with EasySep™ Column-Free Cell Separation Technology
视频
How to Isolate Cells with EasySep™ Column-Free Cell Separation Technology
How EasySep™ Magnetic Cell Separation Technology Works: Fast and Easy Cell Isolation
视频
How EasySep™ Magnetic Cell Separation Technology Works: Fast and Easy Cell Isolation
Online Immunology Journal Club: The Potential of Antibody-Producing B Cells for Modelling and Therapy
视频
Online Immunology Journal Club: The Potential of Antibody-Producing B Cells for Modelling and Therapy
Human Immune Cytokines
挂图
Human Immune Cytokines
Streamline Your B Cell Research from Isolation to Expansion
线上讲座
Streamline Your B Cell Research from Isolation to Expansion
Peptide-Assisted Genome Editing of Primary Cells
线上讲座
Peptide-Assisted Genome Editing of Primary Cells

常见问题 (11)

Can EasySep™ be used for either positive or negative selection?

Yes. The EasySep™ kits use either a negative selection approach by targeting and removing unwanted cells or a positive selection approach targeting desired cells. Depletion kits are also available for the removal of cells with a specific undesired marker (e.g. GlyA).

How does the separation work?

Magnetic particles are crosslinked to cells using Tetrameric Antibody Complexes (TAC). When placed in the EasySep™ Magnet, labeled cells migrate to the wall of the tube. The unlabeled cells are then poured off into a separate fraction.

Which columns do I use?

The EasySep™ procedure is column-free. That's right - no columns!

How can I analyze the purity of my enriched sample?

The Product Information Sheet provided with each EasySep™ kit contains detailed staining information.

Can EasySep™ separations be automated?

Yes. RoboSep™, the fully automated cell separator, automates all EasySep™ labeling and cell separation steps.

Can EasySep™ be used to isolate rare cells?

Yes. We recommend a cell concentration of 2x108 cells/mL and a minimum working volume of 100 µL. Samples containing 2x107 cells or fewer should be suspended in 100 µL of buffer.

Are the EasySep™ magnetic particles FACS-compatible?

Yes, the EasySep™ particles are flow cytometry-compatible, as they are very uniform in size and about 5000X smaller than other commercially available magnetic beads used with column-free systems.

Can the EasySep™ magnetic particles be removed after enrichment?

No, but due to the small size of these particles, they will not interfere with downstream applications.

Can I alter the separation time in the magnet?

Yes; however, this may impact the kit's performance. The provided EasySep™ protocols have already been optimized to balance purity, recovery and time spent on the isolation.

For positive selection, can I perform more than 3 separations to increase purity?

Yes, the purity of targeted cells will increase with additional rounds of separations; however, cell recovery will decrease.

How does the binding of the EasySep™ magnetic particle affect the cells? is the function of positively selected cells altered by the bound particles?

Hundreds of publications have used cells selected with EasySep™ positive selection kits for functional studies. Our in-house experiments also confirm that selected cells are not functionally altered by the EasySep™ magnetic particles.

If particle binding is a key concern, we offer two options for negative selection. The EasySep™ negative selection kits can isolate untouched cells with comparable purities, while RosetteSep™ can isolate untouched cells directly from whole blood without using particles or magnets.

文献 (22)

LLT1 and CD161 Expression in Human Germinal Centers Promotes B Cell Activation and CXCR4 Downregulation. Llibre A et al. Journal of Immunology 2016 MAR

Abstract

Germinal centers (GCs) are microanatomical structures critical for the development of high-affinity Abs and B cell memory. They are organized into two zones,light and dark,with coordinated roles,controlled by local signaling. The innate lectin-like transcript 1 (LLT1) is known to be expressed on B cells,but its functional role in the GC reaction has not been explored. In this study,we report high expression of LLT1 on GC-associated B cells,early plasmablasts,and GC-derived lymphomas. LLT1 expression was readily induced via BCR,CD40,and CpG stimulation on B cells. Unexpectedly,we found high expression of the LLT1 ligand,CD161,on follicular dendritic cells. Triggering of LLT1 supported B cell activation,CD83 upregulation,and CXCR4 downregulation. Overall,these data suggest that LLT1-CD161 interactions play a novel and important role in B cell maturation within the GC in humans.
View publication
CD24 Expression and B Cell Maturation Shows a Novel Link With Energy Metabolism: Potential Implications for Patients With Myalgic Encephalomyelitis/Chronic Fatigue Syndrome. F. F. K. Mensah et al. Frontiers in immunology 2018

Abstract

CD24 expression on pro-B cells plays a role in B cell selection and development in the bone marrow. We previously detected higher CD24 expression and frequency within IgD+ na{\{i}}ve and memory B cells in patients with Myalgic Encephalomyelitis/Chronic Fatigue Syndrome (ME/CFS) compared with age-matched healthy controls (HC). Here we investigated the relationship between CD24 expression and B cell maturation. In vitro stimulation of isolated B cells in response to conventional agonists were used to follow the dynamics of CD24 positivity during proliferation and differentiation (or maturation). The relationship between CD24 expression to cycles of proliferation and metabolism in purified B cells from HC was also investigated using phospho-flow (phosphorylation of AMPK-pAMPK) 1proton nuclear magnetic resonance and Mitotracker Far-red (Mitochondrial mass-MM). In vitro in the absence of stimulation there was an increased percentage of CD24+ viable B cells in ME/CFS patients compared to HC (p {\textless} 0.05) following 5 days culture. Following stimulation with B cell agonists percentage of CD24+B cells in both na{\"{i}}ve and memory B cell populations decreased. P {\textless} 0.01). There was a negative relationship between percentage of CD24+B cells with MM (R2 = 0.76; p {\textless} 0.01) which was subsequently lost over sequential cycles of proliferation. There was a significant correlation between CD24 expression on B cells and the usage of glucose and secretion of lactate in vitro. Short term ligation of the B cell receptor with anti-IgM antibody significantly reduced the viability of CD24+ memory B cells compared to those cross-linked by anti-IgD or anti-IgG antibody. A clear difference was found between na{\""{i}}ve and memory B cells with respect to CD24 expression and pAMPK most notably a strong positive association in IgD+IgM+ memory B cells. In vitro findings confirmed dysregulation of CD24-expressing B cells from ME/CFS patients previously suggested by immunophenotype studies of B cells from peripheral blood. CD24-negative B cells underwent productive proliferation whereas CD24+ B cells were either unresponsive or susceptible to cell death upon BCR-engagement alone. We suggest that CD24 expression may reflect variations in energy metabolism on different B cell subsets."""
View publication
BCR-associated factors driving chronic lymphocytic leukemia cells proliferation ex vivo. C. Schleiss et al. Scientific reports 2019 jan

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.
View publication
View All Publications

更多信息

更多信息
物种 人
Magnet Compatibility • EasySep™ Magnet (Catalog #18000) • “The Big Easy” EasySep™ Magnet (Catalog #18001) • Easy 50 EasySep™ Magnet (Catalog #18002) • EasyEights™ EasySep™ Magnet (Catalog #18103) • RoboSep™-S (Catalog #21000) • Easy 250 EasySep™ Magnet (Catalog #100-0821)
样本来源 PBMC, 白细胞单采术样本
Selection Method Negative
标记抗体
  1. 抗人CD3抗体，clone UCHT1
    抗人CD3抗体，clone UCHT1

    小鼠（BALB/c）Monoclonal IgG1抗体，抗人、黑猩猩CD3

  2. 抗人CD45抗体，clone HI30
    抗人CD45抗体，clone HI30

    小鼠Monoclonal IgG1抗体，抗人、黑猩猩CD45

  3. 抗人CD19抗体，clone HIB19
    抗人CD19抗体，clone HIB19

    抗人、黑猩猩CD19的小鼠Monoclonal IgG1抗体

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