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EasySep™人Pan-B细胞富集试剂盒

免疫磁珠负选不带标记的人pan-B细胞
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产品号 #（选择产品）

产品号 #19554_C

免疫磁珠负选不带标记的人pan-B细胞

产品优势

  • 操作简单、快捷，且无需分离柱
  • 纯度高达99%
  • 分选得到的细胞不带标记

产品组分包括

  • EasySep™人Pan-B细胞富集试剂盒（产品号 #19554）
    • EasySep™人Pan-B细胞富集抗体混合物，1 mL
    • EasySep™ D Magnetic Particles磁珠，2 x 1 mL
  • RoboSep™人Pan-B细胞富集试剂盒（产品号 #19554RF）
    • EasySep™人Pan-B细胞富集抗体混合物，1 mL
    • EasySep™ D Magnetic Particles磁珠，2 x 1 mL
    • RoboSep™ 缓冲液（产品号 #20104）
    • RoboSep™过滤吸头（产品号 #20125）
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要查看实验方案所需的所有配套产品，请参阅《实验方案与技术文档》。
  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

通过免疫磁珠负选，从新鲜或冻存的人外周血单个核细胞 (PBMC)或裂解的白细胞单采术样本中分选出高度纯化的B细胞（包括浆细胞）。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，非目的细胞通过抗体复合物与磁珠标记。表达以下标记的非目的细胞将被去除：CD2、CD3、CD14、CD16、CD36、CD42b、CD56、CD66b、CD123及glyA。使用EasySep™磁极吸附后，通过简单地将目的细胞倾倒或吸取至一个新的试管中，即可将被磁珠标记的细胞与不带标记的目的细胞分离开来。经磁珠分选后，目的B细胞可立即用于流式细胞术、细胞培养或DNA/RNA提取等下游应用。

如需从正常样本中仅分离CD43阴性B细胞，推荐使用 EasySep™ 人B细胞富集试剂盒（产品号 #19054）。

如需从B细胞白血病或淋巴瘤患者或B细胞可能表达CD43、CD36和/或CD123的其他疾病患者的外周血或其他组织中富集B细胞，建议使用不去除CD43的EasySep™

人B细胞富集试剂盒 II（产品号 #17963）。

了解更多EasySep™免疫磁珠技术的工作原理，或者如何通过RoboSep™实现全自动化免疫磁珠细胞分选。探索为您的实验流程优化的更多产品，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）
• RoboSep™-S（产品号 #21000）
 
分类
细胞分选试剂盒
 
细胞类型
B 细胞，血浆
 
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
免疫
 

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实验数据

FACS Profile Results With EasySep™ Human Pan-B Cell Enrichment Kit

Figure 1. Typical EasySep™ Human Pan-B Cell Enrichment Profile

Starting with nucleated cells, the pan-B cell [Lineage (CD4, CD8, CD14, CD16, CD56) negative, CD19+ and CD19-CD43+] content of the enriched fraction typically ranges from 90 - 99%.

Expansion and Maturation of Human B Cells with ImmunoCult™ Human B Cell Expansion Kit

Figure 2. Expansion and Maturation of Human B Cells with ImmunoCult™ Human B Cell Expansion Kit

B cells isolated from human peripheral blood mononuclear cells (PBMCs) using EasySep™ Human Pan-B Cell Enrichment Kit (Catalog #19554) were seeded at 1 x 10⁵ cells/well in 24-well tissue culture plates with ImmunoCult™-ACF Human B Cell Expansion Supplement and ImmunoCult™-XF B Cell Base Medium, included in the ImmunoCult™ Human B Cell Expansion Kit (Catalog #100-0645). The cells were passaged every 3 - 4 days.

(A) Fold expansion of viable cells is shown for n = 12 donors, with bars representing the mean and 95% confidence level (range 38- to 1190-fold at Day 14 ± 1 day).

(B) Expression of CD138 and CD20 was analyzed by flow cytometry at each timepoint (data represent % positive viable cells; mean ± 1 SD). The observed changes indicate maturation of B cells to plasma cells/blasts.

Light Microscopy Image of Cultured Human B Cells

Figure 3. Light Microscopy Image of Cultured Human B Cells

B cells isolated from human PBMCs using EasySep™ Human Pan-B Cell Enrichment Kit (Catalog #19554) were seeded at 1 x 10⁵ cells/well in a 24-well tissue culture plate and cultured with the ImmunoCult™ Human B Cell Expansion Kit (Catalog #100-0646). The cells were passaged on Day 4 after seeding and imaged at 40X magnification on Day 6.

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
Product Information Sheet
Product Name
RoboSep™ Human Pan-B Cell Enrichment Kit
Catalog #
19554RF
Lot #
All
Language
English
Document Type
Product Information Sheet
Product Name
EasySep™ Human Pan-B Cell Enrichment Kit
Catalog #
19554
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Product Name
RoboSep™ Human Pan-B Cell Enrichment Kit
Catalog #
19554RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Product Name
RoboSep™ Human Pan-B Cell Enrichment Kit
Catalog #
19554RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 3
Product Name
RoboSep™ Human Pan-B Cell Enrichment Kit
Catalog #
19554RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Product Name
EasySep™ Human Pan-B Cell Enrichment Kit
Catalog #
19554
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Product Name
EasySep™ Human Pan-B Cell Enrichment Kit
Catalog #
19554
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 (9)

Cell Enrichment Prior to Cell Sorting
Brochure
Cell Enrichment Prior to Cell Sorting
EasySep™ Cell Separation Technology
Brochure
EasySep™ Cell Separation Technology
Tools for Your Immunology Research
Brochure
Tools for Your Immunology Research
Human B Cell Isolation Product Selection
Brochure
Human B Cell Isolation Product Selection
Human Immune Cytokines
Wallchart
Human Immune Cytokines
Frequencies of Human Cell Types in Blood-Related Sources
Wallchart
Frequencies of Human Cell Types in Blood-Related Sources
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
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

常见问题 (12)

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.

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.

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.

Which cell separation kits are compatible with the "Easy 50" EASYSEP™ magnet?

At present, the "Easy 50" EasySep™ magnet is only compatible with EasySep™ kits for human cell separation.

T Cells: 19051 (T Cells), 19052 (CD4 T cells), 19157 (Memory CD4 T Cells), 19053 (CD8 T Cells), 19159 (Memory CD8 T Cells - please contact Tech Support)

B Cells: 19054 (B Cells), 19254 (Naïve B cells)

Other Cell Types: 19055 (NK Cells), 19058 (Monocytes without CD16 depletion), 19059 (Monocytes), 19062 (Plasmacytoid DCs), 19251 (pan-DCs)

For HLA Analysis: 19951HLA (T Cells from whole blood), 19954HLA (B Cells from whole blood), 19961HLA (Total lymphocytes from whole blood)

文献 (9)

Expression and Site-Specific Biotinylation of Human Cytosolic 5′-Nucleotidase 1A in Escherichia coli Methods and Protocols 2025 Jun

Abstract

Autoantibodies targeting cytosolic 5′-nucleotidase 1A (cN1A) are found in several autoimmune diseases, including inclusion body myositis (IBM), Sjögren’s syndrome, and systemic lupus erythematosus. While they have diagnostic relevance for IBM, little is known about the autoreactive B cells that produce these antibodies. To address this, we developed a robust protocol for the expression and site-specific biotinylation of recombinant human cN1A in Escherichia coli. The resulting antigen is suitable for generating double-labelled fluorescent baits for the isolation and characterisation of cN1A-specific B cells by flow cytometry. Site-specific biotinylation was achieved using the AviTag and BirA ligase, preserving the protein’s structure and immunoreactivity. Western blot analysis confirmed that the biotinylated cN1A was recognised by both human and rabbit anti-cN1A antibodies. Compared to conventional chemical biotinylation, this strategy minimises structural alterations that may affect antigen recognition. This approach provides a reliable method for producing biotinylated antigens for use in immunological assays. While demonstrated here for cN1A, the protocol can be adapted for other autoantigens to support studies of antigen-specific B cells in autoimmune diseases.
View publication
Full-length single-cell BCR sequencing paired with RNA sequencing reveals convergent responses to pneumococcal vaccination Communications Biology 2024 Sep

Abstract

Single-cell RNA sequencing (scRNA-seq) can resolve transcriptional features from individual cells, but scRNA-seq techniques capable of resolving the variable regions of B cell receptors (BCRs) remain limited, especially from widely-used 3′-barcoded libraries. Here, we report a method that can recover paired, full-length variable region sequences of BCRs from 3′-barcoded scRNA-seq libraries. We first verify this method (B3E-seq) can produce accurate, full-length BCR sequences. We then apply this method to profile B cell responses elicited against the capsular polysaccharide of Streptococcus pneumoniae serotype 3 (ST3) by glycoconjugate vaccines in five infant rhesus macaques. We identify BCR features associated with specificity for the ST3 antigen which are present in multiple vaccinated monkeys, indicating a convergent response to vaccination. These results demonstrate the utility of our method to resolve key features of the B cell repertoire and profile antigen-specific responses elicited by vaccination. A method that recovers full-length, paired heavy- and light-chain variable regions of B cell receptor transcripts from 3’barcoded scRNA-seq libraries reveals a convergent response to pneumococcus vaccination in rhesus macaques.
View publication
Preclinical specificity & activity of a fully human 41BB-expressing anti-CD19 CART- therapy for treatment-resistant autoimmune disease Molecular Therapy. Methods & Clinical Development 2024 May

Abstract

Over 4% of the global population is estimated to live with autoimmune disease, necessitating immunosuppressive treatment that is often chronic, not curative, and carries associated risks. B cells have emerged as key players in disease pathogenesis, as evidenced by partial responsiveness to B cell depletion by antibody-based therapies. However, these treatments often have transient effects due to incomplete depletion of tissue-resident B cells. Chimeric antigen receptor (CAR) T cells targeting B cells have demonstrated efficacy in refractory systemic lupus erythematosus. To this end, we developed an anti-CD19 CAR T cell product candidate, CABA-201, containing a clinically evaluated fully human CD19 binder (IC78) with a 4-1BB costimulatory domain and CD3 zeta stimulation domain for treatment refractory autoimmune disease. Here, we demonstrate specific cytotoxic activity of CABA-201 against CD19+ Nalm6 cells with no off-target effects on primary human cells. Novel examination of CABA-201 generated from primary T cells from multiple patients with autoimmune disease displayed robust CAR surface expression and effective elimination of the intended target autologous CD19+ B cells in vitro. Together, these findings support the tolerability and activity of CABA-201 for clinical development in patients with autoimmune disease. Graphical abstract Basu and colleagues show CABA-201, a B cell-targeting CAR T cell product with a fully human CD19 binder and 4-1BB costimulatory domain, can precisely eliminate autoimmune patient B cells without off-target deleterious effects, demonstrating its ability as a robust therapeutic for B cell-driven autoimmune disorders.
View publication
View All Publications

更多信息

更多信息
物种 人类
Magnet Compatibility • EasySep™ Magnet (Catalog #18000) • “The Big Easy” EasySep™ Magnet (Catalog #18001) • Easy 50 EasySep™ Magnet (Catalog #18002) • RoboSep™-S (Catalog #21000)
样本来源 PBMC, 白细胞单采术样本
Selection Method Negative
标记抗体
  1. 抗人CD16抗体，clone 3G8
    抗人CD16抗体，clone 3G8

    小鼠抗人、恒河猴、食蟹猴CD16 Monoclonal IgG1抗体

  2. 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

  3. 抗人CD4抗体，clone OKT4
    抗人CD4抗体，clone OKT4

    小鼠Monoclonal IgG2b抗体，抗人、恒河猴、食蟹猴CD4

  4. 抗人CD43抗体，clone CD43-10G7
    抗人CD43抗体，clone CD43-10G7

    小鼠Monoclonal IgG1抗体，抗人CD43

  5. 抗人CD56抗体，clone HCD56
    抗人CD56抗体，clone HCD56

    小鼠Monoclonal IgG1抗体，抗人CD56 （NCAM）

  6. 抗人CD8a抗体，clone RPA-T8
    抗人CD8a抗体，clone RPA-T8

    小鼠Monoclonal IgG1抗体，抗人、恒河猴、食蟹猴CD8a

  7. 抗人CD14抗体，clone M5E2
    抗人CD14抗体，clone M5E2

    抗人、恒河猴、食蟹猴CD14的小鼠Monoclonal IgG2a抗体

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