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⁻）

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¥11,818.00

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）

立即询价

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。

实验数据

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.

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。

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)

Shortened Intervals during Heterologous Boosting Preserve Memory CD8 T Cell Function but Compromise Longevity. Thompson EA et al. Journal of Immunology 2016 APR

Abstract

Developing vaccine strategies to generate high numbers of Ag-specific CD8 T cells may be necessary for protection against recalcitrant pathogens. Heterologous prime-boost-boost immunization has been shown to result in large quantities of functional memory CD8 T cells with protective capacities and long-term stability. Completing the serial immunization steps for heterologous prime-boost-boost can be lengthy,leaving the host vulnerable for an extensive period of time during the vaccination process. We show in this study that shortening the intervals between boosting events to 2 wk results in high numbers of functional and protective Ag-specific CD8 T cells. This protection is comparable to that achieved with long-term boosting intervals. Short-boosted Ag-specific CD8 T cells display a canonical memory T cell signature associated with long-lived memory and have identical proliferative potential to long-boosted T cells Both populations robustly respond to antigenic re-exposure. Despite this,short-boosted Ag-specific CD8 T cells continue to contract gradually over time,which correlates to metabolic differences between short- and long-boosted CD8 T cells at early memory time points. Our studies indicate that shortening the interval between boosts can yield abundant,functional Ag-specific CD8 T cells that are poised for immediate protection; however,this is at the expense of forming stable long-term memory.

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Enhancing Lentiviral and Alpharetroviral Transduction of Human Hematopoietic Stem Cells for Clinical Application. J. W. Schott et al. Molecular therapy. Methods {\&} clinical development 2019 sep

Abstract

Ex vivo retroviral gene transfer into CD34+ hematopoietic stem and progenitor cells (HSPCs) has demonstrated remarkable clinical success in gene therapy for monogenic hematopoietic disorders. However,little attention has been paid to enhancement of culture and transduction conditions to achieve reliable effects across patient and disease contexts and to maximize potential vector usage and reduce treatment cost. We systematically tested three HSPC culture media manufactured to cGMP and eight previously described transduction enhancers (TEs) to develop a state-of-the-art clinically applicable protocol. Six TEs enhanced lentiviral (LV) and five TEs facilitated alpharetroviral (ARV) CD34+ HSPC transduction when used alone. Combinatorial TE application tested with LV vectors yielded more potent effects,with up to a 5.6-fold increase in total expression of a reporter gene and up to a 3.8-fold increase in VCN. Application of one of the most promising combinations,the poloxamer LentiBOOST and protamine sulfate,for GMP-compliant manufacturing of a clinical-grade advanced therapy medicinal product (ATMP) increased total VCN by over 6-fold,with no major changes in global gene expression profiles or inadvertent loss of CD34+CD90+ HSPC populations. Application of these defined culture and transduction conditions is likely to significantly improve ex vivo gene therapy manufacturing protocols for HSPCs and downstream clinical efficacy.

View publication

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.

View publication

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更多信息

更多信息
物种	人
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

EasySep™人Naïve B细胞富集试剂盒

产品号 #19254

产品号 #19254RF

产品号 #（选择产品）

产品号 #19254_C

产品优势

产品组分包括

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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Scientists Helping Scientists™

EasySep™人Naïve B细胞富集试剂盒

产品号 #19254

产品号 #19254RF

产品号 #（选择产品）

产品号 #19254_C

产品优势

产品组分包括

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

技术资料 (10)

常见问题 (11)

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

How does the separation work?

Which columns do I use?

How can I analyze the purity of my enriched sample?

Can EasySep™ separations be automated?

Can EasySep™ be used to isolate rare cells?

Are the EasySep™ magnetic particles FACS-compatible?

Can the EasySep™ magnetic particles be removed after enrichment?

Can I alter the separation time in the magnet?

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

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

文献 (6)

Abstract

Abstract

Abstract

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