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EasySep™人NK细胞分选试剂盒

免疫磁珠负选不带标记的人NK细胞
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产品号 #（选择产品）

产品号 #17955_C

免疫磁珠负选不带标记的人NK细胞

产品优势

  • 操作简单、快捷，且无需分离柱
  • 纯度高达94%
  • 分选得到的细胞不带标记

产品组分包括

  • EasySep™人NK细胞分选试剂盒（产品号 #17955）
    • EasySep™人NK细胞分选抗体混合物，1 x 1 mL
    • EasySep™ Dextran RapidSpheres™磁珠，1 x 1 mL
  • EasySep™人NK细胞分选试剂盒（产品号 #100-0960）
    • EasySep™人NK细胞分选抗体混合物，1 x 10 mL（产品号 #300-0475）
    • EasySep™ Dextran RapidSpheres™磁珠，1 x 1 mL（产品号 #300-0380）
  • RoboSep™人NK细胞分选试剂盒（产品号 #17955RF）
    • EasySep™人NK细胞分选抗体混合物，1 mL
    • EasySep™ Dextran RapidSpheres™磁珠，1 mL
    • RoboSep™ 缓冲液（产品号 #20104）
    • RoboSep™过滤吸头（产品号 #20125）
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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》。
  1. EasySep™磁极
    EasySep™磁极
  2. EasySep™缓冲液
    EasySep™缓冲液

What Our Scientist Says

We know that cell isolation is only part of your workflow. We designed this kit to isolate NK cells in as little as 8 minutes, so you can get to your downstream experiments as soon as possible.

Manreet ChehalScientist
Manreet Chehal Scientist, Scientist
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
相关产品

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（PBMC）或洗涤的白细胞单采术样本中分选出不带磁珠标记和高纯度的自然杀伤（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，非目的细胞通过抗体复合物与磁珠被标记。以下非目标细胞将被去除：粒细胞、T细胞、B细胞、单核细胞、树突状细胞、红细胞。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8分钟的分选后，目的NK细胞可立即用于流式细胞术、培养或DNA/RNA提取等下游应用。

该产品可替代EasySep™人NK细胞富集试剂盒（产品号 #19055）以进行更快的NK细胞负选。

针对包含CD36+和HLA-DR+新生亚群的NK细胞分选，我们推荐EasySep™人Pan-NK细胞分选试剂盒（产品号 #100-1580），短至8分钟即可完成含新生亚群的NK细胞分选。

如需从白细胞单采术样本中大规模分选人NK细胞，请参见大包装（1x10^10 个细胞）的试剂盒（产品号 #100-0960）。

深入了解EasySep™免疫磁珠分选技术的原理，或探索如何通过RoboSep™实现全自动免疫磁珠细胞分选。您亦可选择通过EasySep™人NK细胞分选试剂盒分选的即用型、符合伦理来源的新鲜原代人外周血NK细胞。探索更多为您实验流程优化的其它产品，包括培养基、补充剂、抗体等。

磁极兼容性
• EasySep™磁极（产品号 #18000）
• “The Big Easy” EasySep™磁极（产品号 #18001）
• Easy 50 EasySep™磁极（产品号 #18002）
• EasyEights™ EasySep™磁极（产品号 #18103）
• RoboSep™-S（产品号 #21000）
• Easy 250 EasySep™磁极（产品号 #100-0821）
 
分类
细胞分选试剂盒
 
细胞类型
NK 细胞
 
种属
人
 
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
嵌合体，免疫
 

查看更多 显示更少

实验数据

Separation of natural killer cells using EasySep™ Human NK Cell Isolation Kit

Figure 1. Typical EasySep™ Human NK Cell Isolation Profile

Starting with human PBMCs, the NK cell (CD3-CD56+) content of the isolated fraction is typically 85.0 ± 8.0% (mean ± SD). In the above example, the final purities of the start and isolated fractions are 5.9% and 86.7%, respectively.

Cell Frequency, Fold Expansion, and Phenotyping of Natural Killer (NK) Cells Cultured Using the ImmunoCult™ NK Cell Expansion Kit

Figure 2. CD56+CD3− NK Cells Expand Over 14 Days in Feeder- and Serum-Free Culture Conditions

Human CD56+CD3− NK cells isolated using EasySep™ Human NK Cell Isolation Kit (Catalog #17955) were cultured using ImmunoCult™ NK Cell Expansion Kit (Catalog #100-0711) for 14 days. Cells were harvested and analyzed for expression of characteristic NK cell markers, including CD56, CD3, CD16, CD94, KIR, NKG2D, NKp46, NKp30, and NKp44 by flow cytometry. Staining for killer cell immunoglobulin-like receptor (KIR) molecules was performed using two different antibody clones, HP-MA4 and 180704, which recognize distinct KIR molecules. Dead cells were excluded by light-scatter profile and DRAQ7™ staining. (A - H) Representative flow cytometry plots. (I) The average frequencies of viable CD56+CD3− and CD56+CD16+ NK cells on Day 14 were 87 ± 1% and 75 ± 2%, respectively. The average fold expansion of CD56+CD3− cells was 89 ± 17. Results shown represent mean ± SEM (n = 34).

Cytotoxicity of Expanded Natural Killer (NK) Cells Co-Cultured with K562 Cells

Figure 3. Expanded NK Cells Are Functional, Killing K562 Cells in Co-Culture

CD56+CD3− NK cells isolated using EasySep™ Human NK Cell Isolation Kit (Catalog #17955) were expanded using ImmunoCult™ NK Cell Expansion Kit (Catalog #100-0711) and co-cultured with Incucyte® Cytolight Rapid Dye-labeled K562 cells at a 1:1 ratio of NK:K562 cells at 37°C for 4 hours. Incucyte® Caspase-3/7 Dye, a caspase-inducible dye, was added to the co-culture to detect caspase-induced apoptosis of the K562 cells. Images were obtained every hour using the Incucyte® imaging system and then analyzed to determine % killing (# apoptotic K562 cells ÷ # total labeled K562 cells). After 4 hours, an average of 48 ± 2.4% K562 cells were killed (n = 9). Data represent mean ± SEM.

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
产品说明书
Product Name
RoboSep™ 人NK细胞分选试剂盒
Catalog #
17955RF
Lot #
All
Language
中文
Document Type
产品说明书
Product Name
EasySep™人NK细胞分选试剂盒
Catalog #
17955
Lot #
All
Language
中文
Document Type
Product Information Sheet - with Easy 250 EasySep™ Magnet
Product Name
EasySep™ Human NK Cell Isolation Kit
Catalog #
100-0960
Lot #
All
Language
English
Document Type
Product Information Sheet
Product Name
RoboSep™ Human NK Cell Isolation Kit
Catalog #
17955RF
Lot #
All
Language
English
Document Type
Product Information Sheet
Product Name
EasySep™ Human NK Cell Isolation Kit
Catalog #
17955
Lot #
All
Language
English
Document Type
Safety Data Sheet
Product Name
EasySep™ Human NK Cell Isolation Kit
Catalog #
100-0960
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Product Name
RoboSep™ Human NK Cell Isolation Kit
Catalog #
17955RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Product Name
RoboSep™ Human NK Cell Isolation Kit
Catalog #
17955RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 3
Product Name
RoboSep™ Human NK Cell Isolation Kit
Catalog #
17955RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Product Name
EasySep™ Human NK Cell Isolation Kit
Catalog #
17955
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Product Name
EasySep™ Human NK Cell Isolation Kit
Catalog #
17955
Lot #
All
Language
English
Document Type
Safety Data Sheet 3
Product Name
EasySep™ Human NK Cell Isolation Kit
Catalog #
17955
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

相关材料与文献

技术资料 (18)

Natural Killer Cells
挂图
Natural Killer Cells
The Immune Response to HIV Poster
挂图
The Immune Response to HIV Poster
Human NK Cell Product Workflow
产品手册
Human NK Cell Product Workflow
EasySep™细胞分选技术 - 更高效地分离细胞
产品手册
EasySep™细胞分选技术 - 更高效地分离细胞
Isolate Human Immune Cells
产品手册
Isolate Human Immune Cells
细胞分选相关工具
产品手册
细胞分选相关工具
T细胞治疗研究
产品手册
T细胞治疗研究
CRISPR-Cas9 Genome Editing of Human NK Cells
实验方案
CRISPR-Cas9 Genome Editing of Human NK Cells
How to Measure NK Cell-Mediated Antibody-Dependent Cellular Cytotoxicity Using Flow Cytometry-Based Assays
实验方案
How to Measure NK Cell-Mediated Antibody-Dependent Cellular Cytotoxicity Using Flow Cytometry-Based Assays
How to Measure NK Cell Killing Activity with a Standardized Potency Assay
实验方案
How to Measure NK Cell Killing Activity with a Standardized Potency Assay
Isolate Cells with a Simple Pour-Off: EasySep™ Cell Separation Technology
视频
Isolate Cells with a Simple Pour-Off: EasySep™ Cell Separation Technology
How to Isolate PBMCs from Whole Blood Using Density Gradient Centrifugation (Ficoll™ or Lymphoprep™)
视频
How to Isolate PBMCs from Whole Blood Using Density Gradient Centrifugation (Ficoll™ or Lymphoprep™)
Simultaneous Cell Isolation from Multiple Samples Using the EasyEights™ EasySep™ Magnet
视频
Simultaneous Cell Isolation from Multiple Samples Using the EasyEights™ EasySep™ Magnet
How to Isolate Cells with EasySep™ Column-Free Cell Separation Technology
视频
How to Isolate Cells with EasySep™ Column-Free Cell Separation Technology
How EasySep™ Magnetic Cell Separation Technology Works: Fast and Easy Cell Isolation
视频
How EasySep™ Magnetic Cell Separation Technology Works: Fast and Easy Cell Isolation
How to Isolate Untouched Human NK Cells and Tips for High Recovery
视频
How to Isolate Untouched Human NK Cells and Tips for High Recovery
T Cell Differentiation and Cancer Immunity
视频
T Cell Differentiation and Cancer Immunity
Human Immune Cytokines
挂图
Human Immune Cytokines

常见问题

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 (27)

Stepwise phosphorylation of p65 promotes NF-κB activation and NK cell responses during target cell recognition. Kwon H-J et al. Nature communications 2016

Abstract

NF-κB is a key transcription factor that dictates the outcome of diverse immune responses. How NF-κB is regulated by multiple activating receptors that are engaged during natural killer (NK)-target cell contact remains undefined. Here we show that sole engagement of NKG2D,2B4 or DNAM-1 is insufficient for NF-κB activation. Rather,cooperation between these receptors is required at the level of Vav1 for synergistic NF-κB activation. Vav1-dependent synergistic signalling requires a separate PI3K-Akt signal,primarily mediated by NKG2D or DNAM-1,for optimal p65 phosphorylation and NF-κB activation. Vav1 controls downstream p65 phosphorylation and NF-κB activation. Synergistic signalling is defective in X-linked lymphoproliferative disease (XLP1) NK cells entailing 2B4 dysfunction and required for p65 phosphorylation by PI3K-Akt signal,suggesting stepwise signalling checkpoint for NF-κB activation. Thus,our study provides a framework explaining how signals from different activating receptors are coordinated to determine specificity and magnitude of NF-κB activation and NK cell responses.
View publication
Dose intensification of TRAIL-inducing ONC201 inhibits metastasis and promotes intratumoral NK cell recruitment. J. Wagner et al. The Journal of clinical investigation 2018 JUN

Abstract

ONC201 is a first-in-class,orally active antitumor agent that upregulates cytotoxic TRAIL pathway signaling in cancer cells. ONC201 has demonstrated safety and preliminary efficacy in a first-in-human trial in which patients were dosed every 3 weeks. We hypothesized that dose intensification of ONC201 may impact antitumor efficacy. We discovered that ONC201 exerts dose- and schedule-dependent effects on tumor progression and cell death signaling in vivo. With dose intensification,we note a potent anti-metastasis effect and inhibition of cancer cell migration and invasion. Our preclinical results prompted a change in ONC201 dosing in all open clinical trials. We observed accumulation of activated NK+ and CD3+ cells within ONC201-treated tumors and that NK cell depletion inhibits ONC201 efficacy in vivo,including against TRAIL/ONC201-resistant Bax-/- tumors. Immunocompetent NCR1-GFP mice,in which NK cells express GFP,demonstrated GFP+ NK cell infiltration of syngeneic MC38 colorectal tumors. Activation of primary human NK cells and increased degranulation occurred in response to ONC201. Coculture experiments identified a role for TRAIL in human NK-mediated antitumor cytotoxicity. Preclinical results indicate the potential utility for ONC201 plus anti-PD-1 therapy. We observed an increase in activated TRAIL-secreting NK cells in the peripheral blood of patients after ONC201 treatment. The results offer what we believe to be a unique pathway of immune stimulation for cancer therapy.
View publication
Human antibodies targeting Zika virus NS1 provide protection against disease in a mouse model. M. J. Bailey et al. Nature communications 2018 NOV

Abstract

Zika virus is a mosquito-borne flavivirus closely related to dengue virus that can cause severe disease in humans,including microcephaly in newborns and Guillain-Barr{\'{e}} syndrome in adults. Specific treatments and vaccines for Zika virus are not currently available. Here,we isolate and characterize four monoclonal antibodies (mAbs) from an infected patient that target the non-structural protein NS1. We show that while these antibodies are non-neutralizing,NS1-specific mAbs can engage Fc$\gamma$R without inducing antibody dependent enhancement (ADE) of infection in vitro. Moreover,we demonstrate that mAb AA12 has protective efficacy against lethal challenges of African and Asian lineage strains of Zika virus in Stat2-/- mice. Protection is Fc-dependent,as a mutated antibody unable to activate known Fc effector functions or complement is not protective in vivo. This study highlights the importance of the ZIKV NS1 protein as a potential vaccine antigen.
View publication
View All Publications

更多信息

更多信息
物种 人
Magnet Compatibility • EasySep™ Magnet (Catalog #18000) • “The Big Easy” EasySep™ Magnet (Catalog #18001) • Easy 50 EasySep™ Magnet (Catalog #18002) • EasyEights™ EasySep™ Magnet (Catalog #18103) • RoboSep™-S (Catalog #21000) • Easy 250 EasySep™ Magnet (Catalog #100-0821)
样本来源 PBMC, 白细胞单采术样本
Selection Method Negative
标记抗体
  1. 抗人CD45抗体，clone HI30
    抗人CD45抗体，clone HI30

    小鼠Monoclonal IgG1抗体，抗人、黑猩猩CD45

  2. 抗人CD56抗体，clone HCD56
    抗人CD56抗体，clone HCD56

    小鼠Monoclonal IgG1抗体，抗人CD56 （NCAM）

  3. 抗人CD3抗体，clone UCHT1
    抗人CD3抗体，clone UCHT1

    小鼠（BALB/c）Monoclonal IgG1抗体，抗人、黑猩猩CD3

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