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EasySep™人Naïve B细胞分选试剂盒

免疫磁珠负选未标记的人Naïve B细胞
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产品号 #（选择产品）

产品号 #17254_C

免疫磁珠负选未标记的人Naïve B细胞

产品优势

  • 操作简单、快捷，且无需分离柱
  • 纯度高达98%，回收率高
  • 分选得到的细胞不带标记

产品组分包括

  • EasySep™人Naïve B细胞分选试剂盒（产品号 #17254）
    • EasySep™ 人Naïve 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™ 人Naïve B细胞分选试剂盒（产品号 #17254RF）
    • EasySep™ 人Naïve 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）
main product photo
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
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
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总览

使用 EasySep™人Naïve B细胞分选试剂盒，可轻松高效地从新鲜或冻存的人外周血单个核细胞（PBMCs）或洗涤后的白细胞单采样本中，通过免疫磁珠负选分选法获得高纯度人人Naïve B细胞（CD3-CD19+CD27-）。EasySep™技术结合单克隆抗体的特异性和无柱磁分选系统的简便性，已在发表的研究中广泛应用超过20年。

在该EasySep™负选流程中，非目标细胞用抗体复合物与磁珠标记。表达以下标志物的非目标细胞将被特异性去除：CD2、CD14、CD16、CD27、CD36、CD43及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，接着简单地将目的细胞倾倒或吸至一个新的试管中。磁珠分选最快仅需9分钟，所得Naïve B细胞可立即用于流式细胞术、培养或DNA/RNA提取等下游应用。

本产品可替代EasySep™人 Naïve B细胞富集试剂盒 (产品号 #19254) 以进行更快地细胞分选。

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。探索更多为您实验流程优化的其它产品，包括培养基、补充剂、抗体等。

磁体兼容性
• 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)
 
亚型
细胞分选试剂盒
 
细胞类型
B 细胞
 
种属
人
 
样本来源
Leukapheresis，PBMC
 
筛选方法
Negative
 
应用
细胞分选
 
品牌
EasySep，RoboSep
 
研究领域
免疫
 

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Data Figures

Typical EasySep™ Human Naïve B Cell Isolation Profile

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

Starting with human PBMCs, the naïve B cell (CD3-CD19+CD27-) content of the isolated fraction is typically 94.9 ± 2.2% (mean ± SD). In the above example, the purities of the start and final isolated fractions are 7.1% and 97.7%, respectively.

Protocols and Documentation

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 Naïve B Cell Isolation Kit
Catalog #
17254
Lot #
Purchase date 2020-04-07 and later
Language
English
Document Type
Product Information Sheet 1
Product Name
RoboSep™ Human Naïve B Cell Isolation Kit
Catalog #
17254RF
Lot #
All other lots
Language
English
Document Type
Product Information Sheet 2
Product Name
RoboSep™ Human Naïve B Cell Isolation Kit
Catalog #
17254RF
Lot #
Purchase date 2020-04-07 and later
Language
English
Document Type
Safety Data Sheet 1
Product Name
EasySep™ Human Naïve B Cell Isolation Kit
Catalog #
17254
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Product Name
EasySep™ Human Naïve B Cell Isolation Kit
Catalog #
17254
Lot #
All
Language
English
Document Type
Safety Data Sheet 3
Product Name
EasySep™ Human Naïve B Cell Isolation Kit
Catalog #
17254
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Product Name
RoboSep™ Human Naïve B Cell Isolation Kit
Catalog #
17254RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Product Name
RoboSep™ Human Naïve B Cell Isolation Kit
Catalog #
17254RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 3
Product Name
RoboSep™ Human Naïve B Cell Isolation Kit
Catalog #
17254RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 4
Product Name
RoboSep™ Human Naïve B Cell Isolation Kit
Catalog #
17254RF
Lot #
All
Language
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.

Research Area
Workflow Stages

Resources and Publications

Educational Materials (11)

EasySep™ Cell Separation Technology
Brochure
EasySep™ Cell Separation Technology
Human B Cell Isolation Product Selection
Brochure
Human B Cell Isolation Product Selection
Tools for Your Immunology Research
Brochure
Tools for Your Immunology Research
Isolate Human Immune Cells
Brochure
Isolate Human Immune Cells
Frequencies of Human Cell Types in Blood-Related Sources
Wallchart
Frequencies of Human Cell Types in Blood-Related Sources
Human Immune Cytokines
Wallchart
Human Immune Cytokines
How to Isolate Cells with EasySep™ Column-Free Cell Separation Technology
1:23
Video
How to Isolate Cells with EasySep™ Column-Free Cell Separation Technology
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
Simultaneous Cell Isolation from Multiple Samples Using the EasyEights™ EasySep™ Magnet
0:57
Video
Simultaneous Cell Isolation from Multiple Samples Using the EasyEights™ EasySep™ Magnet
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...

Frequently Asked Questions

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.

Publications (2)

Inhibition of PI3K p110$\delta$ activity reduces IgE production in IL-4 and anti-CD40 stimulated human B cell cultures. A. Cutrina-Pons et al. Immunology 2023 dec

Abstract

Phosphoinositide 3-kinase (PI3K) p110$\delta$ signalling negatively regulates the production of mouse IgE. However, there are disparities between the mouse and human IgE biology, and the role of PI3K p110$\delta$ in the production of human IgE is yet to be determined. To investigate the effect of PI3K p110$\delta$ inhibition in the production of human IgE we isolated human B cells from tonsil tissue and stimulated them with IL-4 and anti-CD40 antibody to induce class switching to IgE and IgG1 in the presence or absence of IC87114, a small molecule inhibitor of PI3K p110$\delta$. Using FACS, RT-PCR and ELISA we examined the effect of PI3K p110$\delta$ inhibition on IgE production and determined the mechanisms involved. Unlike in mice, we observed that PI3K p110$\delta$ inhibition significantly reduces the number of IgE+ switched cells and the amounts of secreted IgE in IL4 and anti-CD40 cultures. However, the number of IgG1+ cells and secreted IgG1 were largely unaffected by PI3K p110$\delta$ inhibition. The expression levels of AID, $\epsilon$ and $\gamma$1 germinal transcripts or other factors involved in the regulation of CSR to IgE and IgG1 were also unaffected by IC87114. However, we found that IC87114 significantly decreases the proliferation of tonsil B cells stimulated with IL-4 and anti-CD40, specifically reducing the frequency of cells that had undergone 4 divisions or more. In addition, PI3K p110$\delta$ inhibition reduced the levels of IRF4 expression in IgE+ germinal centre-like B cells leading to a block in plasma cell differentiation. In conclusion, PI3K p110$\delta$ signalling is required for the production of human IgE, which makes it a pharmacological target for the treatment of allergic disease.
View publication
Trypanosoma cruzi Induces Regulatory B Cell Alterations in Patients With Chronic Chagas Disease. M. C. Girard et al. Frontiers in cellular and infection microbiology 2021

Abstract

The clinical evolution of patients with chronic Chagas disease (CCD) is mainly associated with an excessive inflammation and a defective immunomodulatory profile caused by the interaction between T. cruzi and the host. Regulatory B (Breg) cells exert immune suppression mostly through IL-10 production (B10 cells), but also through IL-10-independent mechanisms. Previously, we demonstrated that CCD patients with cardiomyopathy show changes in the ex vivo Breg cell phenotypic distribution although maintain IL-10 production capacity. Here, we sought to identify potential alterations on Breg cells upon in vitro stimulation. Isolated B cells from CCD patients with or without cardiomyopathy and non-infected (NI) donors were stimulated with T. cruzi lysate or CpG + CD40L, and characterized by flow cytometry based on the expression of CD24, CD27, CD38, and the regulatory molecules IL-10 and PD-L1. IL-10 and IL-17 secretion in the supernatant of B cells was evaluated by ELISA. Data showed that T. cruzi stimulation diminished the expression of CD24 and CD38 on CD27- B cells while reducing the percentage of CD24high inside CD27+ B cells. Furthermore, T. cruzi induced a regulatory B cell phenotype by increasing B10 cells and IL-10 secretion in all the groups. The innate-like B10 cells expansion observed in patients with cardiomyopathy would be associated with CD27- B10 cell subsets, while no predominant phenotype was found in the other groups. Patients with cardiomyopathy also displayed higher IL-17 secretion levels in T. cruzi-activated B cells. CpG + CD40L stimulation revealed that B cells from CCD patients and NI donors had the same ability to differentiate into B10 cells and secrete IL-10 in vitro. Additionally, CCD patients showed an increased frequency of CD24-CD27- B cells and a reduction in the percentage of CD24highCD27+ Breg cells, which appeared to be inversely correlated with the presence of T. cruzi DNA in blood. Finally, CCD patients exhibited a higher frequency of PD-L1+ B cells in T. cruzi-stimulated samples, suggesting that IL-10-independent mechanisms could also be tangled in the control of inflammation. Altogether, our results provide evidence about the potential role of Breg cells in the immune response developed against T. cruzi and its contribution to chronic Chagas cardiomyopathy.
View publication
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更多信息

更多信息
Species Human
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)
Sample Source Leukapheresis, PBMC
Selection Method Negative
标记抗体
  1. 抗人CD45抗体，克隆HI30
    抗人CD45抗体，克隆HI30

    小鼠单克隆IgG1抗体，抗人、黑猩猩CD45

  2. 抗人CD3抗体，克隆UCHT1
    抗人CD3抗体，克隆UCHT1

    小鼠（BALB/c）单克隆IgG1抗体，抗人、黑猩猩CD3

  3. 抗小鼠CD27抗体，克隆LG.3A10
    抗小鼠CD27抗体，克隆LG.3A10

    抗人、小鼠、大鼠CD27的仓鼠（亚美尼亚）单克隆IgG1抗体

  4. 抗人CD19抗体，克隆HIB19
    抗人CD19抗体，克隆HIB19

    抗人、黑猩猩CD19的小鼠单克隆IgG1抗体

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