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EasySep™人Naïve B细胞富集试剂盒

对来源于外周血单个核细胞或白细胞分离样本进行免疫磁珠分离,获得未标记的人初始 B 细胞(CD19⁺CD27⁻)

产品号 #(选择产品)

产品号 #19254_C

免疫磁珠负选

产品优势

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

产品组分包括

  • EasySep™人Naïve B细胞富集试剂盒(产品号 #19254)
    • EasySep™人Naïve B细胞富集抗体混合物,1 mL
    • EasySep™ D Magnetic Particles磁珠,5 x 1 mL
  • EasySep™人Naïve B细胞富集试剂盒(含过滤吸头)(产品号 #19254RF)
    • EasySep™人Naïve B细胞富集抗体混合物,1 mL
    • EasySep™ D Magnetic Particles磁珠,5 x 1 mL
    • RoboSep™ 缓冲液(产品号 #20104)
    • RoboSep™过滤吸头(产品号 #20125)
专为您的实验方案打造的产品
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》

总览

使用 EasySep™ 人初始 B 细胞富集试剂盒,可通过免疫磁性负选法从新鲜或冻存的人外周血单个核细胞(PBMCs)或白细胞单采样本中轻松高效地分离高纯度的初始 B 细胞(CD19⁺CD27⁻)。EasySep™ 技术在已发表的研究中被广泛应用超过 20 年,结合了单克隆抗体的特异性与无柱磁分选系统的简便性。

在此 EasySep™ 负选步骤中,非目的细胞被抗体复合物和磁性颗粒标记。以下标志物阳性的细胞将被去除:CD2、CD3、CD14、CD16、CD27、CD36、CD43、CD56、CD66b 和 GlyA。经 EasySep™ 磁极分离后,带有磁珠标记的非目的细胞被去除,未标记的目的初始 B 细胞通过倾倒或移液方式收集。磁性分离完成后,初始 B 细胞可直接用于流式细胞术、培养或 DNA/RNA 提取等下游应用。

了解更多关于 EasySep™ 免疫磁性技术的工作原理,或了解如何使用 RoboSep™ 实现全自动化免疫磁性细胞分离。也可选择使用 EasySep™ 人初始 B 细胞富集试剂盒分离的、来源合伦理的现成冷冻人外周血初始 B 细胞(CD19⁺CD27⁻)。探索更多产品,包括培养基、补充物、抗体等,以优化您的实验流程。

 

磁极兼容性
• EasySep™磁极(产品号 #18000)
• “The Big Easy” EasySep™磁极(产品号 #18001)
• EasyPlate™ EasySep™磁极(产品号 #18102)
• 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
 
种属

 
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
 

实验数据

FACS Profile Results With EasySep™ Human Naïve B Cell Enrichment Kit

Figure 1. FACS Profile Results With EasySep™ Human Naïve B Cell Enrichment Kit

Starting with fresh mononuclear cells, the naïve B cell (CD19+CD27-) content of the isolated fraction typically ranges from 92 - 98%. In the above example, the purities of the start and isolated fractions are 4.3% and 97.5%, 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
Catalog #
19254RF
Lot #
All
Language
English
Catalog #
19254
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Catalog #
19254RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Catalog #
19254RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 3
Catalog #
19254RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Catalog #
19254
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Catalog #
19254
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。

相关材料与文献

技术资料 (10)

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 (6)

Targeting RAB7 in human B lymphoma by a small molecule inhibitor arrests tumor cell growth M. Fernandez et al. Frontiers in Oncology 2025 Sep

Abstract

RAB7, encoded by RAB7A in humans and Rab7 in mice, is a small GTPase that catalyzes endosome maturation. It mediates NF-κB activation through the assembly of intracellular membrane signalosomes in stimulated normal B cells and plays a B cell-intrinsic role in the antibody response in mice. Here we show RAB7A transcripts are expressed in primary diffuse large B-cell lymphomas (DLBCLs), and that RAB7 protein expression is heightened in activated human tonsil B cells as well as in DLBCL and Burkitt lymphoma cell lines. Treating these cell lines with CID1067700, a selective small-molecule RAB7 inhibitor, results in a dose-dependent decrease in cell growth, associated with impaired proliferation and survival. CID1067700 also suppressed tumor development from Daudi cells, a Burkitt lymphoma cell line, in Foxn1nu/nu nude mice. The inhibitory effect of CID1067700 on Daudi cell growth in vitro is further enhanced by methyl-β-cyclodextrin, which disrupts plasma membrane lipid rafts, and by FX1, a BCL6 inhibitor. These findings, together with the unfavorable prognosis of DLBCL patients showing high RAB7A expression, suggest that targeting RAB7 is a promising therapeutic approach for mature B cell-derived lymphomas.
CRL3-SPOP ubiquitin ligase complex suppresses the growth of diffuse large B-cell lymphoma by negatively regulating the MyD88/NF-$\kappa$B signaling. X. Jin et al. Leukemia 2020 may

Abstract

Recurrent oncogenic mutations of MyD88 have been identified in a variety of lymphoid malignancies. Gain-of-function mutations of MyD88 constitutively activate downstream NF-$\kappa$B signaling pathways, resulting in increased cellular proliferation and survival. However, whether MyD88 activity can be aberrantly regulated in MyD88-wild-type lymphoid malignancies remains poorly understood. SPOP is an adaptor protein of CUL3-based E3 ubiquitin ligase complex and frequently mutated genes in prostate and endometrial cancers. In this study, we reveal that SPOP binds to and induces the nondegradative ubiquitination of MyD88 by recognizing an atypical SPOP-binding motif in MyD88. This modification blocks Myddosome assembly and downstream NF-$\kappa$B activation. SPOP is mutated in a subset of lymphoid malignancies, including diffuse large B-cell lymphoma (DLBCL). Lymphoid malignancies-associated SPOP mutants exhibited impaired binding to MyD88 and suppression of NF-$\kappa$B activation. The DLBCL-associated, SPOP-binding defective mutants of MyD88 escaped from SPOP-mediated ubiquitination, and their effect on NF-$\kappa$B activation is stronger than that of wild-type MyD88. Moreover, SPOP suppresses DLBCL cell growth in vitro and tumor xenograft in vivo by inhibiting the MyD88/NF-$\kappa$B signaling. Therefore, SPOP acts as a tumor suppressor in DLBCL. Mutations in the SPOP-MyD88 binding interface may disrupt the SPOP-MyD88 regulatory axis and promote aberrant MyD88/NF-$\kappa$B activation and cell growth in DLCBL.
B cell-intrinsic epigenetic modulation of antibody responses by dietary fiber-derived short-chain fatty acids. H. N. Sanchez et al. Nature communications 2020

Abstract

Short-chain fatty acids (SCFAs) butyrate and propionate are metabolites from dietary fiber's fermentation by gut microbiota that can affect differentiation or functions of T cells, macrophages and dendritic cells. We show here that at low doses these SCFAs directly impact B cell intrinsic functions to moderately enhance class-switch DNA recombination (CSR), while decreasing at higher doses over a broad physiological range, AID and Blimp1 expression, CSR, somatic hypermutation and plasma cell differentiation. In human and mouse B cells, butyrate and propionate decrease B cell Aicda and Prdm1 by upregulating select miRNAs that target Aicda and Prdm1 mRNA-3'UTRs through inhibition of histone deacetylation (HDAC) of those miRNA host genes. By acting as HDAC inhibitors, not as energy substrates or through GPR-engagement signaling in these B cell-intrinsic processes, these SCFAs impair intestinal and systemic T-dependent and T-independent antibody responses. Their epigenetic impact on B cells extends to inhibition of autoantibody production and autoimmunity in mouse lupus models.

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) • EasyPlate™ EasySep™ Magnet (Catalog 18102) • 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

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