总览

使用 EasySep™ 小鼠 CD117 (cKIT) 正选试剂盒，可通过免疫磁珠正选法从小鼠骨髓或其他组织的单细胞悬液中高效分离高纯度的 CD117⁺（cKIT⁺）细胞。EasySep™ 技术在发表研究中已被广泛使用超过 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™ 正选步骤中， CD117⁺ 目的细胞首先使用 EasySep™ PE 正选试剂混合物标记，该混合物含有结合 CD117 的藻红蛋白（PE）标记抗体。随后，这些 PE 标记的抗体被能够同时识别 PE 和右旋糖的抗体复合物及磁性颗粒靶向识别。经 EasySep™ 磁极分离后，不需要的细胞被倒出， CD117⁺ 目的细胞保留在试管中。磁性细胞分离后，CD117⁺ 细胞已带有 PE 标记，可直接用于流式细胞术、细胞培养及 DNA/RNA 提取等下游应用。

了解更多关于 EasySep™ 免疫磁性细胞分选技术的工作原理，或了解如何使用 RoboSep™ 实现全自动化免疫磁性细胞分离。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）
• RoboSep™-S（产品号 #21000）

分类
细胞分选试剂盒

细胞类型
造血干/祖细胞

种属
小鼠

样本来源
骨髓

分选方法
正选

应用
细胞分选

品牌
EasySep，RoboSep

研究领域
免疫，干细胞生物学

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实验数据

Figure 1. FACS Profile Results with EasySep™ Mouse CD117 (c-KIT) Positive Selection Kit

Starting with mouse bone marrow, the CD117+ cell content of the selected cells typically ranges from 88 - 95%.

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

EasySep™ Mouse CD117 (cKIT) Positive Selection Kit

Catalog #

18757

Lot #

All

Language

English

Document Type

Product Information Sheet

Product Name

RoboSep™ Mouse CD117 (cKIT) Positive Selection Kit with Filter Tips

Catalog #

18757RF

Lot #

All

Language

English

Document Type

Safety Data Sheet 1

Product Name

EasySep™ Mouse CD117 (cKIT) Positive Selection Kit

Catalog #

18757

Lot #

All

Language

English

Document Type

Safety Data Sheet 2

Product Name

EasySep™ Mouse CD117 (cKIT) Positive Selection Kit

Catalog #

18757

Lot #

All

Language

English

Document Type

Safety Data Sheet 3

Product Name

EasySep™ Mouse CD117 (cKIT) Positive Selection Kit

Catalog #

18757

Lot #

All

Language

English

Document Type

Safety Data Sheet 1

Product Name

RoboSep™ Mouse CD117 (cKIT) Positive Selection Kit with Filter Tips

Catalog #

18757RF

Lot #

All

Language

English

Document Type

Safety Data Sheet 2

Product Name

RoboSep™ Mouse CD117 (cKIT) Positive Selection Kit with Filter Tips

Catalog #

18757RF

Lot #

All

Language

English

Document Type

Safety Data Sheet 3

Product Name

RoboSep™ Mouse CD117 (cKIT) Positive Selection Kit with Filter Tips

Catalog #

18757RF

Lot #

All

Language

English

Document Type

Safety Data Sheet 4

Product Name

RoboSep™ Mouse CD117 (cKIT) Positive Selection Kit with Filter Tips

Catalog #

18757RF

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

Workflow Stages for Hematopoietic Stem and Progenitor Cell Research

Cell Sourcing and Isolation

Expansion and Differentiation

Analysis

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.

TGF$\beta$R-SMAD3 Signaling Induces Resistance to PARP Inhibitors in the Bone Marrow Microenvironment. B. V. Le et al. Cell reports 2020 oct

Abstract

Synthetic lethality triggered by PARP inhibitor (PARPi) yields promising therapeutic results. Unfortunately, tumor cells acquire PARPi resistance, which is usually associated with the restoration of homologous recombination, loss of PARP1 expression, and/or loss of DNA double-strand break (DSB) end resection regulation. Here, we identify a constitutive mechanism of resistance to PARPi. We report that the bone marrow microenvironment (BMM) facilitates DSB repair activity in leukemia cells to protect them against PARPi-mediated synthetic lethality. This effect depends on the hypoxia-induced overexpression of transforming growth factor beta receptor (TGF$\beta$R) kinase on malignant cells, which is activated by bone marrow stromal cells-derived transforming growth factor beta 1 (TGF-$\beta$1). Genetic and/or pharmacological targeting of the TGF-$\beta$1-TGF$\beta$R kinase axis results in the restoration of the sensitivity of malignant cells to PARPi in BMM and prolongs the survival of leukemia-bearing mice. Our finding may lead to the therapeutic application of the TGF$\beta$R inhibitor in patients receiving PARPis.

View publication

Hematopoietic Stem/Progenitor Cell Dependent Participation of Innate Lymphoid Cells in Low-Intensity Sterile Inflammation. S. Korniotis et al. Frontiers in immunology 2018

Abstract

Hematopoietic stem/progenitor cells (HSPC) are characterized by their unique capacities of self-renewal and multi-differentiation potential. This second property makes them able to adapt their differentiation profile depending on the local environment they reach. Taking advantage of an animal model of peritonitis, induced by injection of the TLR-2 ligand, zymosan, we sought to study the relationship between bone marrow-derived hematopoietic stem/progenitor cells (BM-HSPCs) and innate lymphoid cells (ILCs) regarding their emergence and differentiation at the site of inflammation. Our results demonstrate that the strength of the inflammatory signals affects the capacity of BM-derived HSPCs to migrate and give rise in situ to ILCs. Both low- and high-dose of zymosan injections trigger the appearance of mature ILCs in the peritoneal cavity where the inflammation occurs. Herein, we show that only in low-dose injected mice, the recovered ILCs are dependent on an in situ differentiation of BM-derived HSPCs and/or ILC2 precursors (ILC2P) wherein high-dose, the stronger inflammatory environment seems to be able to induce the emergence of ILCs independently of BM-derived HSPCs. We suggest that a relationship between HSPCs and ILCs seems to be affected by the strength of the inflammatory stimuli opening new perspectives in the manipulation of these early hematopoietic cells.

View publication

Treatment of ongoing autoimmune encephalomyelitis with activated B-cell progenitors maturing into regulatory B cells. Korniotis S et al. Nature communications 2016

Abstract

The influence of signals perceived by immature B cells during their development in bone marrow on their subsequent functions as mature cells are poorly defined. Here, we show that bone marrow cells transiently stimulated in vivo or in vitro through the Toll-like receptor 9 generate proB cells (CpG-proBs) that interrupt experimental autoimmune encephalomyelitis (EAE) when transferred at the onset of clinical symptoms. Protection requires differentiation of CpG-proBs into mature B cells that home to reactive lymph nodes, where they trap T cells by releasing the CCR7 ligand, CCL19, and to inflamed central nervous system, where they locally limit immunopathogenesis through interleukin-10 production, thereby cooperatively inhibiting ongoing EAE. These data demonstrate that a transient inflammation at the environment, where proB cells develop, is sufficient to confer regulatory functions onto their mature B-cell progeny. In addition, these properties of CpG-proBs open interesting perspectives for cell therapy of autoimmune diseases.

View publication

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EasySep™小鼠CD117（cKIT）正选试剂盒

产品号 #18757RF

产品号 #18757

产品号 #（选择产品）

产品号 #18757_C

产品优势

产品组分包括

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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™

物种	小鼠
Magnet Compatibility	• EasySep™ Magnet (Catalog #18000) • “The Big Easy” EasySep™ Magnet (Catalog #18001) • RoboSep™-S (Catalog #21000)
样本来源	骨髓
Selection Method	Positive

EasySep™小鼠CD117（cKIT）正选试剂盒

产品号 #18757RF

产品号 #18757

产品号 #（选择产品）

产品号 #18757_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 (6)

常见问题

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 (8)

Abstract

Abstract

Abstract

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