总览

使用EasySep™小鼠NK细胞分选试剂盒，可通过免疫磁珠负选，轻松高效地从小鼠脾脏单细胞悬液及其他组织样本中分离高纯度的小鼠自然杀伤（NK）细胞（CD3-CD49b+）。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。表达以下标志物的非目的细胞将被靶向去除：CD4、CD5、Ly6G、CD8a、F4/80、CD3e、CD19、CD24、TCRgd及7-4。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。在短至25分钟的磁珠分选后，目的NK细胞可立即用于流式细胞术、细胞培养、基于细胞的实验等下游应用。

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。探索更多为您的实验流程优化的产品，包括培养基、补充剂、抗体等。

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。表达以下标志物的非目的细胞将被靶向去除：CD4、CD5、Ly6G、CD8a、F4/80、CD3e、CD19、CD24、TCRgd及7-4。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。在短至25分钟的磁珠分选后，目的NK细胞可立即用于流式细胞术、细胞培养、基于细胞的实验等下游应用。

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。探索更多为您的实验流程优化的产品，包括培养基、补充剂、抗体等。

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。表达以下标志物的非目的细胞将被靶向去除：CD4、CD5、Ly6G、CD8a、F4/80、CD3e、CD19、CD24、TCRgd及7-4。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。在短至25分钟的磁珠分选后，目的NK细胞可立即用于流式细胞术、细胞培养、基于细胞的实验等下游应用。

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。探索更多为您的实验流程优化的产品，包括培养基、补充剂、抗体等。

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。表达以下标志物的非目的细胞将被靶向去除：CD4、CD5、Ly6G、CD8a、F4/80、CD3e、CD19、CD24、TCRgd及7-4。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。在短至25分钟的磁珠分选后，目的NK细胞可立即用于流式细胞术、细胞培养、基于细胞的实验等下游应用。

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。探索更多为您的实验流程优化的产品，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) • EasyEights™ EasySep™ Magnet (Catalog #18103) • RoboSep™-S (Catalog #21000)

亚型
细胞分选试剂盒

细胞类型
NK 细胞

种属
小鼠

样本来源
Spleen

筛选方法
Negative

应用
细胞分选

品牌
EasySep，RoboSep

研究领域
免疫

查看更多显示更少

实验数据

Figure 1. Typical EasySep™ Mouse NK Cell Isolation Profile

Starting with mouse splenocytes, the NK cell (CD3-CD49b+) content of the isolated fraction typically ranges from 67 - 89%.

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™ Mouse NK Cell Isolation Kit

Catalog #

19855RF

Lot #

1000100904 and higher

Language

English

Document Type

Product Information Sheet

Product Name

EasySep™ Mouse NK Cell Isolation Kit

Catalog #

19855

Lot #

1000100904 and higher

Language

English

Document Type

Safety Data Sheet 1

Product Name

RoboSep™ Mouse NK Cell Isolation Kit

Catalog #

19855RF

Lot #

All

Language

English

Document Type

Safety Data Sheet 2

Product Name

RoboSep™ Mouse NK Cell Isolation Kit

Catalog #

19855RF

Lot #

All

Language

English

Document Type

Safety Data Sheet 3

Product Name

RoboSep™ Mouse NK Cell Isolation Kit

Catalog #

19855RF

Lot #

All

Language

English

Document Type

Safety Data Sheet 1

Product Name

EasySep™ Mouse NK Cell Isolation Kit

Catalog #

19855

Lot #

All

Language

English

Document Type

Safety Data Sheet 2

Product Name

EasySep™ Mouse NK Cell Isolation Kit

Catalog #

19855

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 Natural Killer Cell Research

Cell Sourcing

Cell Isolation

Cell Expansion and Differentiation

Cell Characterization

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.

Targeting CISH enhances natural cytotoxicity receptor signaling and reduces NK cell exhaustion to improve solid tumor immunity. P.-L. Bernard et al. Journal for immunotherapy of cancer 2022 may

Abstract

BACKGROUND The success and limitations of current immunotherapies have pushed research toward the development of alternative approaches and the possibility to manipulate other cytotoxic immune cells such as natural killer (NK) cells. Here, we targeted an intracellular inhibiting protein 'cytokine inducible SH2-containing protein' (CISH) in NK cells to evaluate the impact on their functions and antitumor properties. METHODS To further understand CISH functions in NK cells, we developed a conditional Cish-deficient mouse model in NK cells (Cishfl/flNcr1Ki/+ ). NK cells cytokine expression, signaling and cytotoxicity has been evaluated in vitro. Using intravenous injection of B16F10 melanoma cell line and EO711 triple negative breast cancer cell line, metastasis evaluation was performed. Then, orthotopic implantation of breast tumors was performed and tumor growth was followed using bioluminescence. Infiltration and phenotype of NK cells in the tumor was evaluated. Finally, we targeted CISH in human NK-92 or primary NK cells, using a technology combining the CRISPR(i)-dCas9 tool with a new lentiviral pseudotype. We then tested human NK cells functions. RESULTS In Cishfl/flNcr1Ki/+ mice, we detected no developmental or homeostatic difference in NK cells. Global gene expression of Cishfl/flNcr1Ki/+ NK cells compared with Cish+/+Ncr1Ki/+ NK cells revealed upregulation of pathways and genes associated with NK cell cycling and activation. We show that CISH does not only regulate interleukin-15 (IL-15) signaling pathways but also natural cytotoxicity receptors (NCR) pathways, triggering CISH protein expression. Primed Cishfl/flNcr1Ki/+ NK cells display increased activation upon NCR stimulation. Cishfl/flNcr1Ki/+ NK cells display lower activation thresholds and Cishfl/flNcr1Ki/+ mice are more resistant to tumor metastasis and to primary breast cancer growth. CISH deletion favors NK cell accumulation to the primary tumor, optimizes NK cell killing properties and decreases TIGIT immune checkpoint receptor expression, limiting NK cell exhaustion. Finally, using CRISPRi, we then targeted CISH in human NK-92 or primary NK cells. In human NK cells, CISH deletion also favors NCR signaling and antitumor functions. CONCLUSION This study represents a crucial step in the mechanistic understanding and safety of Cish targeting to unleash NK cell antitumor function in solid tumors. Our results validate CISH as an emerging therapeutic target to enhance NK cell immunotherapy.

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A vaccine targeting resistant tumours by dual T cell plus NK cell attack. S. Badrinath et al. Nature 2022 jun

Abstract

Most cancer vaccines target peptide antigens, necessitating personalization owing to the vast inter-individual diversity in major histocompatibility complex (MHC) molecules that present peptides to T cells. Furthermore, tumours frequently escape T cell-mediated immunity through mechanisms that interfere with peptide presentation1. Here we report a cancer vaccine that induces a coordinated attack by diverse T cell and natural killer (NK) cell populations. The vaccine targets the MICA and MICB (MICA/B) stress proteins expressed by many human cancers as a result of DNA damage2. MICA/B serve as ligands for the activating NKG2D receptor on T cells and NK cells, but tumours evade immune recognition by proteolytic MICA/B cleavage3,4. Vaccine-induced antibodies increase the density of MICA/B proteins on the surface of tumour cells by inhibiting proteolytic shedding, enhance presentation of tumour antigens by dendritic cells to T cells and augment the cytotoxic function of NK cells. Notably, this vaccine maintains efficacy against MHC class I-deficient tumours resistant to cytotoxic T cells through the coordinated action of NK cells and CD4+ T cells. The vaccine is also efficacious in a clinically important setting: immunization following surgical removal of primary, highly metastatic tumours inhibits the later outgrowth of metastases. This vaccine design enables protective immunity even against tumours with common escape mutations.

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The Combination of Radiotherapy and Complement C3a Inhibition Potentiates Natural Killer cell Functions Against Pancreatic Cancer. Q. H. Sodji et al. Cancer research communications 2022 jul

Abstract

Pancreatic cancer is one of the deadliest cancers, against which current immunotherapy strategies are not effective. Herein, we analyzed the immune cell composition of the tumor microenvironment of pancreatic cancer samples in The Cancer Genome Atlas and found that the presence of intratumoral NK cells correlates with survival. Subsequent analysis also indicated that NK cell exclusion from the microenvironment is found in a high percentage of clinical pancreatic cancers and in preclinical models of pancreatic cancer. Mechanistically, NK cell exclusion is regulated in part by complement C3a and its receptor signaling. Inhibition of the C3a receptor enhances NK cell infiltration in syngeneic mouse models of pancreatic cancer resulting in tumor growth delay. However, tumor growth inhibition mediated by NK cells is not sufficient alone for complete tumor regression, but is potentiated when combined with radiation therapy. Our findings indicate that although C3a inhibition is a promising approach to enhance NK cell-based immunotherapy against pancreatic cancer, its combination with radiation therapy hold greater therapeutic benefit.

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EasySep™小鼠NK细胞分选试剂盒

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产品号 #19855_C

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Scientists Helping Scientists™

种属	Mouse
Magnet Compatibility	• EasySep™ Magnet (Catalog #18000) • “The Big Easy” EasySep™ Magnet (Catalog #18001) • EasyEights™ EasySep™ Magnet (Catalog #18103) • RoboSep™-S (Catalog #21000)
样本来源	Spleen
Selection Method	Negative

EasySep™小鼠NK细胞分选试剂盒

产品号 #（选择产品）

产品号 #19855_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 (7)

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 (13)

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