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EasySep™ Direct人中性粒细胞分选试剂盒

直接从全血中免疫磁珠分选中性粒细胞
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产品号 #（选择产品）

产品号 #19666_C

直接从全血中免疫磁珠分选中性粒细胞

产品优势

  • 99.9%红细胞去除率，无需密度梯度离心、沉降或裂解
  • 分选获得的细胞纯度高达99%
  • 操作简单、快捷，且无需分离柱
  • 分选得到的细胞不带标记

产品组分包括

  • EasySep™ Direct人中性粒细胞分选试剂盒(产品号 #19666)    
    • EasySep™ Direct人中性粒细胞分选抗体混合物, 2 x 2.5 mL
    • EasySep™ Direct RapidSpheres™ 磁珠, 4 x 2.5 mL
  • RoboSep™人中性粒细胞分选试剂盒 (产品号 #100-0404)    
    • EasySep™ Direct人中性粒细胞分选抗体混合物, 2 x 2.5 mL
    • EasySep™ Direct RapidSpheres™ 磁珠, 4 x 2.5 mL
    • RoboSep™ 缓冲液（产品号 #20104）
    • RoboSep™过滤吸头（产品号#20125）
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  1. EasySep™磁极
    EasySep™磁极

What Our Scientist Says

RBC lysis and other preprocessing steps can complicate cell isolation procedures. We developed this kit so you can keep things simple and quickly isolate neutrophils directly from whole blood.

Christine BarrScientist
Christine Barr, 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

使用EasySep™ Direct 人中性粒细胞分选试剂盒 ，可轻松高效地从人全血样本中通过免疫磁珠负选获得高纯度人中性粒细胞。EasySep™技术结合单克隆抗体的特异性和免磁柱系统的简便性，已在发表的研究中广泛应用超过20年。

在该EasySep™负 选流程中，非目的细胞会被抗体复合物和EasySep™ Direct RapidSpheres™ 磁珠标记。以下非目的细胞会被特异性去除：嗜碱性粒细胞、嗜酸性粒细胞、T细胞、B细胞、单核细胞、NK细胞和红系细胞。通过EasySep™磁极将被磁珠标记的细胞与未被标记的目的细胞分离，目的细胞可通过倾倒或者移液吸取到新试管中，分选后的细胞 可直接用于流式细胞术、细胞培养或DNA/RNA提取等下游应用。

该产品可替代EasySep™人中性粒细胞分选试剂盒 (产品号 #19257) 以实现更快地细胞分选。

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，包括细胞鉴定 、冷冻保存等相关试剂 。

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）
 
分类
细胞分选试剂盒
 
细胞类型
粒细胞及其亚群
 
种属
人
 
样本来源
全血
 
分选方法
负选
 
应用
细胞分选
 
品牌
EasySep，RoboSep
 
研究领域
药物发现和毒性检测，免疫学，传染病
 

查看更多 显示更少

实验数据

Starting with Human Whole Blood from Normal Healthy Donors, the Typical Neutrophil (CD66b+CD16+) Content of the Non-lysed Final Isolated Fraction Is 97.3 ± 1.4% (Gated on CD45) or 94.0 ± 3.7% (Not Gated on CD45)

Figure 1. Typical EasySep™ Direct Human Neutrophil Isolation Profile

Starting with human whole blood from normal healthy donors, the typical neutrophil (CD66b+CD16+) content of the non-lysed final isolated fraction Is 97.3 ± 1.4% (gated on CD45) or 94.0 ± 3.7% (not gated on CD45). In the example above, the neutrophil (CD66b+CD16+) content of the lysed whole blood start sample and the non-lysed final isolated fraction is 50.6% and 97.2% (gated on CD45), respectively, or 43.6% and 95.9% (not gated on CD45), respectively. The starting frequency of neutrophils in the non-lysed whole blood start sample above is 0.04% (data not shown).

Typical EasySep™ Direct Protocol

Figure 2. Typical EasySep™ Direct Protocol

Use of Highly Enriched Lymphocytes Isolated with EasySep™ Direct Improves DSA Detection Compared to Whole Leukocyte Cell Preparations

Figure 3. Use of Highly Enriched Lymphocytes Isolated with EasySep™ Direct Improves DSA Detection Compared to Whole Leukocyte Cell Preparations

Lymphocytes (Ly), neutrophils (Nu), and monocytes (Mo) were isolated from volunteer donors (n = 5) using EasySep™ Direct Catalog #19655, #19666, and #19669, respectively. Whole leukocyte (WL) preparations were obtained by adding Ly, Nu, and Mo cells in equal propotions. WL (low lymphocyte purity) and Ly (high ymphocyte purity) preparations were treated with pronase and then used to perform the flow cytometry cross-match (FCXM) assay against negative control sera or several dilutions of positive control sera. The median channel fluorescence shifts (MCFS) were generated by using the negative control sera samples as a baseline. The MCF shifts between WL and Ly were then compared. Each column with error bars represents the mean ± SEM (n = 5 donors). Data kindly provided by Dr. Robert Liwski.

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™ 人中性粒细胞分选试剂盒
Catalog #
100-0404
Lot #
All
Language
中文
Document Type
产品说明书
Product Name
EasySep™ Direct人中性粒细胞分选试剂盒
Catalog #
19666
Lot #
All
Language
中文
Document Type
Product Information Sheet
Product Name
RoboSep™ Human Neutrophil Isolation Kit
Catalog #
100-0404
Lot #
All
Language
English
Document Type
Product Information Sheet
Product Name
EasySep™ Direct Human Neutrophil Isolation Kit
Catalog #
19666
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Product Name
RoboSep™ Human Neutrophil Isolation Kit
Catalog #
100-0404
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Product Name
RoboSep™ Human Neutrophil Isolation Kit
Catalog #
100-0404
Lot #
All
Language
English
Document Type
Safety Data Sheet 3
Product Name
RoboSep™ Human Neutrophil Isolation Kit
Catalog #
100-0404
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Product Name
EasySep™ Direct Human Neutrophil Isolation Kit
Catalog #
19666
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Product Name
EasySep™ Direct Human Neutrophil Isolation Kit
Catalog #
19666
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 (10)

血液相关来源中人细胞类型的比例
挂图
血液相关来源中人细胞类型的比例
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
EasySep™ Direct 仅需20分钟，从全血中分选细胞
产品手册
EasySep™ Direct 仅需20分钟，从全血中分选细胞
Isolation of Untouched Neutrophils Directly from Whole Blood in 20 Minutes
科学海报
Isolation of Untouched Neutrophils Directly from Whole Blood in 20 Minutes
Isolate Cells with a Simple Pour-Off: EasySep™ Cell Separation Technology
视频
Isolate Cells with a Simple Pour-Off: EasySep™ Cell Separation Technology
Cell Isolation Directly from Whole Blood without RBC Lysis or Centrifugation: EasySep™ Direct
视频
Cell Isolation Directly from Whole Blood without RBC Lysis or Centrifugation: EasySep™ Direct
How to Isolate Cells Directly from Whole Blood Using the EasySep™ Purple/Silver Magnets
视频
How to Isolate Cells Directly from Whole Blood Using the EasySep™ Purple/Silver Magnets
How to Isolate Cells Directly from Whole Blood - EasySep™ Direct Protocol, EasyEights Magnet
视频
How to Isolate Cells Directly from Whole Blood - EasySep™ Direct Protocol, EasyEights Magnet
How to Isolate Cells Directly from Whole Blood Using the Easy 50 EasySep™ Magnet: 30-Minute Protocol
视频
How to Isolate Cells Directly from Whole Blood Using the Easy 50 EasySep™ Magnet: 30-Minute Protocol
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 (55)

Neutrophils can disarm NK cell response through cleavage of NKp46. Valayer A et al. Journal of leukocyte biology 2016 SEP

Abstract

Polymorphonuclear neutrophils (PMNs) can contribute to the regulation of the host immune response by crosstalk with innate and adaptive leukocytes,including NK cells. Mechanisms by which this immunoregulation process occurs remain incompletely understood. Here,we focused on the effect of human neutrophil-derived serine proteases on NKp46,a crucial activating receptor expressed on NK cells. We used flow cytometry,Western blotting,and mass spectrometry (MS) analysis to reveal that cathepsin G [CG; and not elastase or proteinase 3 (PR3)] induces a time- and concentration-dependent,down-regulatory effect on NKp46 expression through a restricted proteolytic mechanism. We also used a functional assay to demonstrate that NKp46 cleavage by CG severely impairs NKp46-mediated responses of NK cells,including IFN-γ production and cell degranulation. Importantly,sputa of cystic fibrosis (CF) patients,which have high concentrations of CG,also alter NKp46 on NK cells. Hence,we have identified a new immunoregulatory mechanism of neutrophils that proteolytically disarms NK cell responses.
View publication
Imprime PGG-Mediated Anti-Cancer Immune Activation Requires Immune Complex Formation. A. S. H. Chan et al. PloS one 2016

Abstract

Imprime PGG (Imprime),an intravenously-administered,soluble $\beta$-glucan,has shown compelling efficacy in multiple phase 2 clinical trials with tumor targeting or anti-angiogenic antibodies. Mechanistically,Imprime acts as pathogen-associated molecular pattern (PAMP) directly activating innate immune effector cells,triggering a coordinated anti-cancer immune response. Herein,using whole blood from healthy human subjects,we show that Imprime-induced anti-cancer functionality is dependent on immune complex formation with naturally-occurring,anti-$\beta$ glucan antibodies (ABA). The formation of Imprime-ABA complexes activates complement,primarily via the classical complement pathway,and is opsonized by iC3b. Immune complex binding depends upon Complement Receptor 3 and Fcg Receptor IIa,eliciting phenotypic activation of,and enhanced chemokine production by,neutrophils and monocytes,enabling these effector cells to kill antibody-opsonized tumor cells via the generation of reactive oxygen species and antibody-dependent cellular phagocytosis. Importantly,these innate immune cell changes were not evident in subjects with low ABA levels but could be rescued with exogenous ABA supplementation. Together,these data indicate that pre-existing ABA are essential for Imprime-mediated anti-cancer immune activation and suggest that pre-treatment ABA levels may provide a plausible patient selection biomarker to delineate patients most likely to benefit from Imprime-based therapy.
View publication
Germline-encoded neutralization of a Staphylococcus aureus virulence factor by the human antibody repertoire. Yeung YA et al. Nature communications 2016 NOV

Abstract

Staphylococcus aureus is both an important pathogen and a human commensal. To explore this ambivalent relationship between host and microbe,we analysed the memory humoral response against IsdB,a protein involved in iron acquisition,in four healthy donors. Here we show that in all donors a heavily biased use of two immunoglobulin heavy chain germlines generated high affinity (pM) antibodies that neutralize the two IsdB NEAT domains,IGHV4-39 for NEAT1 and IGHV1-69 for NEAT2. In contrast to the typical antibody/antigen interactions,the binding is primarily driven by the germline-encoded hydrophobic CDRH-2 motifs of IGHV1-69 and IGHV4-39,with a binding mechanism nearly identical for each antibody derived from different donors. Our results suggest that IGHV1-69 and IGHV4-39,while part of the adaptive immune system,may have evolved under selection pressure to encode a binding motif innately capable of recognizing and neutralizing a structurally conserved protein domain involved in pathogen iron acquisition.
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)
样本来源 全血
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CD66b抗体，clone G10F5
    抗人CD66b抗体，clone G10F5

    小鼠Monoclonal IgM抗体，抗人、黑猩猩CD66b

  3. 抗人CD16抗体，clone 3G8
    抗人CD16抗体，clone 3G8

    小鼠抗人、恒河猴、食蟹猴CD16 Monoclonal IgG1抗体

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