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EasySep™小鼠中性粒细胞富集试剂盒

免疫磁珠负选未标记的小鼠中性粒细胞
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产品号 #（选择产品）

产品号 #19762_C

免疫磁珠负选未标记的小鼠中性粒细胞

产品优势

  • 操作简单、快捷，且无需分离柱
  • 纯度高达93.7%（血液）和88.7%（骨髓）。
  • 分选得到的细胞不带标记

产品组分包括

  • EasySep™小鼠中性粒细胞富集试剂盒 （产品号 #19762）
    • EasySep™小鼠中性粒细胞富集抗体混合物，0.5 mL
    • EasySep™生物素分选抗体混合物，1 mL
    • EasySep™ 磁珠，2 x 1 mL
    • EasySep™ 小鼠FcR阻断剂（产品号 #18730），0.2 mL
  • RoboSep™ 小鼠中性粒细胞富集试剂盒（含过滤吸头）（产品号19762RF）
    • EasySep™小鼠中性粒细胞富集抗体混合物，0.5 mL
    • EasySep™生物素分选抗体混合物，1 mL
    • EasySep™ 磁珠，2 x 1 mL
    • EasySep™ 小鼠FcR阻断剂（产品号 #18730），0.2 mL
    • RoboSep™ 缓冲液（产品号 #20104）
    • RoboSep™ 过滤吸头（产品号 #20125）
main product photo
New format, same high quality! You may notice that your kit contents and packaging look slightly different from previous orders. We are currently updating the format of select EasySep™ Mouse kits to include a Mouse FcR blocker instead of Normal Rat Serum. With this change, all components will now be shipped in a single package, while providing the same cell isolation performance as before.
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

EasySep™ 小鼠中性粒细胞富集试剂盒，通过免疫磁珠负选，轻松高效地从小鼠骨髓、外周血及其他组织样本的单细胞悬液中分离高纯度小鼠中性粒细胞（CD11b+Ly6G+）。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。表达以下标志物的非目标细胞将被定向去除：CD4、CD5、Ter119、CD45R、CD49b、F4/80、CD117和CD11c。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，接着简单地将目的细胞倾倒或吸取至一个新的试管中。经磁珠分选后，目的小鼠中性粒细胞即可用于流式细胞术、细胞培养或基于细胞的检测分析等下游应用。

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。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) • RoboSep™-S (Catalog #21000)
 
亚型
细胞分选试剂盒
 
细胞类型
粒细胞及其亚群
 
种属
小鼠
 
样本来源
Bone Marrow，Whole Blood
 
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

Typical EasySep™ Mouse Neutrophil Cell Isolation Profile

Figure 1. Typical EasySep™ Mouse Neutrophil Cell Isolation Profile

Starting with mouse bone marrow or mouse blood, the CD11b+Ly6G+ cell content of the enriched fraction is typically 88.2 ± 3.2% for bone marrow and 88.6 ± 4.9% for blood (mean ± SD) using the purple EasySep™ magnet. In the above example, the purities of the start and final enriched fractions are 46.6% and 89.1% (bone marrow) and 20.1% and 91.5% (blood), respectively.

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 1
Product Name
EasySep™ Mouse Neutrophil Enrichment Kit
Catalog #
19762
Lot #
1000138269 or higher
Language
English
Document Type
Product Information Sheet 2
Product Name
EasySep™ Mouse Neutrophil Enrichment Kit
Catalog #
19762
Lot #
1000138268 or lower
Language
English
Document Type
Product Information Sheet 1
Product Name
RoboSep™ Mouse Neutrophil Enrichment Kit with Filter Tips
Catalog #
19762RF
Lot #
1000138269 or higher
Language
English
Document Type
Product Information Sheet 2
Product Name
RoboSep™ Mouse Neutrophil Enrichment Kit with Filter Tips
Catalog #
19762RF
Lot #
1000138268 or lower
Language
English
Document Type
Safety Data Sheet 1
Product Name
EasySep™ Mouse Neutrophil Enrichment Kit
Catalog #
19762
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Product Name
EasySep™ Mouse Neutrophil Enrichment Kit
Catalog #
19762
Lot #
All
Language
English
Document Type
Safety Data Sheet 3
Product Name
EasySep™ Mouse Neutrophil Enrichment Kit
Catalog #
19762
Lot #
All
Language
English
Document Type
Safety Data Sheet 4
Product Name
EasySep™ Mouse Neutrophil Enrichment Kit
Catalog #
19762
Lot #
All
Language
English
Document Type
Safety Data Sheet 5
Product Name
EasySep™ Mouse Neutrophil Enrichment Kit
Catalog #
19762
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Product Name
RoboSep™ Mouse Neutrophil Enrichment Kit with Filter Tips
Catalog #
19762RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Product Name
RoboSep™ Mouse Neutrophil Enrichment Kit with Filter Tips
Catalog #
19762RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 3
Product Name
RoboSep™ Mouse Neutrophil Enrichment Kit with Filter Tips
Catalog #
19762RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 4
Product Name
RoboSep™ Mouse Neutrophil Enrichment Kit with Filter Tips
Catalog #
19762RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 5
Product Name
RoboSep™ Mouse Neutrophil Enrichment Kit with Filter Tips
Catalog #
19762RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 6
Product Name
RoboSep™ Mouse Neutrophil Enrichment Kit with Filter Tips
Catalog #
19762RF
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

Resources and Publications

Educational Materials (4)

EasySep™ Cell Separation Technology
Brochure
EasySep™ Cell Separation Technology
Human Immune Cytokines
Wallchart
Human Immune Cytokines
Frequencies and Percentages of Mouse Immune Cell Types
Wallchart
Frequencies and Percentages of Mouse Immune Cell Types
Isolation of Neutrophils from Mouse Bone Marrow and Blood Using a Rapid
Scientific Poster
Isolation of Neutrophils from Mouse Bone Marrow and Blood Using a Rapid

Frequently Asked Questions

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.

Publications (28)

Liraglutide enhances the effect of checkpoint blockade in lung and liver cancers through the inhibition of neutrophil extracellular traps. D. Chen et al. FEBS open bio 2024 aug

Abstract

Glucagon-like peptide-1 (GLP-1) regulates glycemic excursions by augmenting insulin production and inhibiting glucagon secretion. Liraglutide, a long-acting GLP-1 analog, can improve glycemic control for treating type 2 diabetes and prevent neutrophil extravasation in inflammation. Here, we explored the role of liraglutide in the development and therapy of murine lung and liver cancers. In this study, liraglutide substantially decreased circulating neutrophil extracellular trap (NET) markers myeloperoxidase, elastase, and dsDNA in Lewis lung cancer (LLC) and Hepa1-6 tumor-bearing mice. Furthermore, liraglutide downregulated NETs and reactive oxygen species (ROS) of neutrophils in the tumor microenvironment. Functionally, in??vitro experiments showed that liraglutide reduced NET formation by inhibiting ROS. In addition, we showed that liraglutide enhanced the anti-tumoral efficiency of programmed cell death-1 (PD-1) inhibition in LLC and Hepa1-6 tumor-bearing C57BL/6 mice. However, the removal of NETs significantly weakened the antitumor efficiency of liraglutide. We further demonstrated that the long-term antitumor CD8+ T cell responses induced by the combination therapy rejected rechallenges by respective tumor cell lines. Taken together, our findings suggest that liraglutide may promote the anti-tumoral efficiency of PD-1 inhibition by reducing NETs in lung and liver cancers.
View publication
The voltage-gated potassium channel KV1.3 regulates neutrophil recruitment during inflammation. R. Immler et al. Cardiovascular research 2022 mar

Abstract

Aims: Neutrophil trafficking within the vasculature strongly relies on intracellular calcium signalling. Sustained Ca2+ influx into the cell requires a compensatory efflux of potassium to maintain membrane potential. Here, we aimed to investigate whether the voltage-gated potassium channel KV1.3 regulates neutrophil function during the acute inflammatory process by affecting sustained Ca2+ signalling. Methods and results: Using in vitro assays and electrophysiological techniques, we show that KV1.3 is functionally expressed in human neutrophils regulating sustained store-operated Ca2+ entry through membrane potential stabilizing K+ efflux. Inhibition of KV1.3 on neutrophils by the specific inhibitor 5-(4-Phenoxybutoxy)psoralen (PAP-1) impaired intracellular Ca2+ signalling, thereby preventing cellular spreading, adhesion strengthening, and appropriate crawling under flow conditions in vitro. Using intravital microscopy, we show that pharmacological blockade or genetic deletion of KV1.3 in mice decreased neutrophil adhesion in a blood flow dependent fashion in inflamed cremaster muscle venules. Furthermore, we identified KV1.3 as a critical component for neutrophil extravasation into the inflamed peritoneal cavity. Finally, we also revealed impaired phagocytosis of Escherichia coli particles by neutrophils in the absence of KV1.3. Conclusion: We show that the voltage-gated potassium channel KV1.3 is critical for Ca2+ signalling and neutrophil trafficking during acute inflammatory processes. Our findings do not only provide evidence for a role of KV1.3 for sustained calcium signalling in neutrophils affecting key functions of these cells, they also open up new therapeutic approaches to treat inflammatory disorders characterized by overwhelming neutrophil infiltration.
View publication
Cure of syngeneic carcinomas with targeted IL-12 through obligate reprogramming of lymphoid and myeloid immunity. Y. Hong et al. JCI insight 2022 mar

Abstract

Therapeutic IL-12 has demonstrated the ability to reduce local immune suppression in preclinical models, but clinical development has been limited by severe inflammation-related adverse events with systemic administration. Here, we show that potent immunologic tumor control of established syngeneic carcinomas can be achieved by i.t. administration of a tumor-targeted IL-12 antibody fusion protein (NHS-rmIL-12) using sufficiently low doses to avoid systemic toxicity. Single-cell transcriptomic analysis and ex vivo functional assays of NHS-rmIL-12-treated tumors revealed reinvigoration and enhanced proliferation of exhausted CD8+ T lymphocytes, induction of Th1 immunity, and a decrease in Treg number and suppressive capacity. Similarly, myeloid cells transitioned toward inflammatory phenotypes and displayed reduced suppressive capacity. Cell type-specific IL-12 receptor-KO BM chimera studies revealed that therapeutic modulation of both lymphoid and myeloid cells is required for maximum treatment effect and tumor cure. Study of single-cell data sets from human head and neck carcinomas revealed IL-12 receptor expression patterns similar to those observed in murine tumors. These results describing the diverse mechanisms underlying tumor-directed IL-12-induced antitumor immunity provide the preclinical rationale for the clinical study of i.t. NHS-IL-12.
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更多信息

更多信息
Species Mouse
Magnet Compatibility • EasySep™ Magnet (Catalog #18000) • “The Big Easy” EasySep™ Magnet (Catalog #18001) • RoboSep™-S (Catalog #21000)
Sample Source Bone Marrow, Whole Blood
Selection Method Negative
标记抗体
  1. 抗小鼠Gr-1抗体，克隆RB6-8C5
    抗小鼠Gr-1抗体，克隆RB6-8C5

    大鼠单克隆IgG2b抗体，抗小鼠Gr-1 （Ly-6G/Ly-6C）

  2. 抗小鼠Ly-6G抗体，克隆1A8
    抗小鼠Ly-6G抗体，克隆1A8

    大鼠单克隆IgG2a抗体，抗小鼠Ly-6G

  3. 抗小鼠CD11b抗体，克隆M1/70
    抗小鼠CD11b抗体，克隆M1/70

    抗人、小鼠、恒河猴CD11b的大鼠单克隆IgG2b抗体

PRODUCTS ARE FOR RESEARCH USE ONLY AND NOT INTENDED FOR HUMAN OR ANIMAL DIAGNOSTIC OR THERAPEUTIC USES UNLESS OTHERWISE STATED. FOR ADDITIONAL INFORMATION ON QUALITY AT STEMCELL, REFER TO WWW.STEMCELL.COM/COMPLIANCE.
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