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EasySep™人NK细胞富集试剂盒

免疫磁珠负选未标记的人NK细胞

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产品号 #（选择产品）

产品号 #19055_C

免疫磁珠负选未标记的人NK细胞

产品优势

操作简单、快捷，且无需分离柱
纯度高达95%
获得的活细胞无标记

产品组分包括

EasySep™ 人NK 细胞富集试剂盒（产品号 #19055）

EasySep™人NK细胞富集抗体混合物, 1 mL
EasySep™ D Magnetic Particles磁珠, 2 x 1 mL

RoboSep™ 人NK细胞富集试剂盒（含过滤吸头）（产品号 #19055RF）

EasySep™人NK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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New look, same high quality and support! You may notice that your instrument or reagent packaging looks slightly different from images displayed on the website, or from previous orders. We are updating our look but rest assured, the products themselves and how you should use them have not changed. Learn more

总览

实验数据

产品说明书及文档

应用领域

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 (PBMCs) 或裂解的白细胞单采术样本中分离出无磁珠标记和高纯度的NK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，非目的细胞被抗体复合物与磁珠标记。以下非目标细胞将被特异性去除：粒细胞、T细胞、B细胞、单核细胞、树突状细胞及红系细胞。通过EasySep™磁极将被磁珠标记的细胞与未被标记的目的人NK细胞分离，接着简单地将目的细胞倾倒或吸取至一个新的试管中。完成磁珠细胞分选后，目的NK细胞即可用于流式细胞术、培养或DNA/RNA提取等下游应用。

如需更快速的细胞分选，我们推荐使用EasySep™ 人NK细胞分选试剂盒（产品号 #17955），仅需8分钟即可完成细胞分选。

了解更多关于免疫磁珠EasySep™技术的工作原理，或如何通过RoboSep™实现免疫磁珠细胞分选全自动化。或选择即用型、符合伦理来源的用EasySep™ 人NK细胞富集试剂盒进行分选的冻存原代人外周血NK细胞。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。

磁体兼容性
• EasySep™ Magnet (Catalog #18000) • “The Big Easy” EasySep™ Magnet (Catalog #18001) • Easy 50 EasySep™ Magnet (Catalog #18002) • EasyPlate™ EasySep™ Magnet (Catalog 18102) • 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
人

样本来源
PBMC

筛选方法
Negative

应用
细胞分选

品牌
EasySep，RoboSep

研究领域
免疫

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Data Figures

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

The NK cell content of the enriched fraction varies, depending on the starting sample. Starting with previously frozen mononuclear cells containing more than 10% NK cells, the NK cell content of the enriched fraction typically ranges from 73% - 95%. Purities may be lower when starting with samples containing less than 10% NK cells.

Protocols and Documentation

Find supporting information and directions for use in the Product Information Sheet or explore additional protocols below.

Document Type

Product Name

Catalog #

Lot #

Language

Document Type

Product Information Sheet

Product Name

RoboSep™ Human NK Cell Enrichment Kit with Filter Tips

Catalog #

19055RF

Lot #

All

Language

English

Document Type

Product Information Sheet

Product Name

EasySep™ Human NK Cell Enrichment Kit

Catalog #

19055

Lot #

All

Language

English

Document Type

Safety Data Sheet 1

Product Name

RoboSep™ Human NK Cell Enrichment Kit with Filter Tips

Catalog #

19055RF

Lot #

All

Language

English

Document Type

Safety Data Sheet 2

Product Name

RoboSep™ Human NK Cell Enrichment Kit with Filter Tips

Catalog #

19055RF

Lot #

All

Language

English

Document Type

Safety Data Sheet 3

Product Name

RoboSep™ Human NK Cell Enrichment Kit with Filter Tips

Catalog #

19055RF

Lot #

All

Language

English

Document Type

Safety Data Sheet 1

Product Name

EasySep™ Human NK Cell Enrichment Kit

Catalog #

19055

Lot #

All

Language

English

Document Type

Safety Data Sheet 2

Product Name

EasySep™ Human NK Cell Enrichment Kit

Catalog #

19055

Lot #

All

Language

English

Applications

This product is designed for use in the following research area(s) as part of the highlighted workflow stage(s). Explore these workflows to learn more about the other products we offer to support each research area.

Research Area

Workflow Stages

Workflow Stages for Natural Killer Cell Research

Cell Sourcing

Cell Isolation

Cell Expansion and Differentiation

Cell Characterization

Resources and Publications

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.

View publication

PRDX-1 Supports the Survival and Antitumor Activity of Primary and CAR-Modified NK Cells under Oxidative Stress. M. Klopotowska et al. Cancer immunology research 2022 feb

Abstract

Oxidative stress, caused by the imbalance between reactive species generation and the dysfunctional capacity of antioxidant defenses, is one of the characteristic features of cancer. Here, we quantified hydrogen peroxide in the tumor microenvironment (TME) and demonstrated that hydrogen peroxide concentrations are elevated in tumor interstitial fluid isolated from murine breast cancers in vivo, when compared with blood or normal subcutaneous fluid. Therefore, we investigated the effects of increased hydrogen peroxide concentration on immune cell functions. NK cells were more susceptible to hydrogen peroxide than T cells or B cells, and by comparing T, B, and NK cells' sensitivities to redox stress and their antioxidant capacities, we identified peroxiredoxin-1 (PRDX1) as a lacking element of NK cells' antioxidative defense. We observed that priming with IL15 protected NK cells' functions in the presence of high hydrogen peroxide and simultaneously upregulated PRDX1 expression. However, the effect of IL15 on PRDX1 expression was transient and strictly dependent on the presence of the cytokine. Therefore, we genetically modified NK cells to stably overexpress PRDX1, which led to increased survival and NK cell activity in redox stress conditions. Finally, we generated PD-L1-CAR NK cells overexpressing PRDX1 that displayed potent antitumor activity against breast cancer cells under oxidative stress. These results demonstrate that hydrogen peroxide, at concentrations detected in the TME, suppresses NK cell function and that genetic modification strategies can improve CAR NK cells' resistance and potency against solid tumors.

View publication

Immunomodulation of T- and NK-cell Responses by a Bispecific Antibody Targeting CD28 Homolog and PD-L1. M. Ramaswamy et al. Cancer immunology research 2022 feb

Abstract

Checkpoint blockade therapies targeting PD-1/PD-L1 and CTLA-4 are clinically successful but also evoke adverse events due to systemic T-cell activation. We engineered a bispecific, mAb targeting CD28 homolog (CD28H), a newly identified B7 family receptor that is constitutively expressed on T and natural killer (NK) cells, with a PD-L1 antibody to potentiate tumor-specific immune responses. The bispecific antibody led to T-cell costimulation, induced NK-cell cytotoxicity of PD-L1-expressing tumor cells, and activated tissue-resident memory CD8+ T cells. Mechanistically, the CD28H agonistic arm of the bispecific antibody reduced PD-L1/PD-1-induced SHP2 phosphorylation while simultaneously augmenting T-cell receptor signaling by activating the MAPK and AKT pathways. This bispecific approach could be used to target multiple immune cells, including CD8+ T cells, tissue-resident memory T cells, and NK cells, in a tumor-specific manner that may lead to induction of durable, therapeutic antitumor responses.

View publication

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EasySep™人NK细胞富集试剂盒

产品号 #（选择产品）

产品号 #19055_C

产品优势

产品组分包括

总览

Data Figures

Protocols and Documentation

Applications

Resources and Publications

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Scientists Helping Scientists™

Species	Human
Magnet Compatibility	• EasySep™ Magnet (Catalog #18000) • “The Big Easy” EasySep™ Magnet (Catalog #18001) • Easy 50 EasySep™ Magnet (Catalog #18002) • EasyPlate™ EasySep™ Magnet (Catalog 18102) • EasyEights™ EasySep™ Magnet (Catalog #18103) • RoboSep™-S (Catalog #21000)
Sample Source	PBMC
Selection Method	Negative

EasySep™人NK细胞富集试剂盒

产品号 #（选择产品）

产品号 #19055_C

产品优势

产品组分包括

总览

Data Figures

Protocols and Documentation

Applications

Resources and Publications

Educational Materials (11)

Frequently Asked Questions

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?

Publications (43)

Abstract

Abstract

Abstract

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