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EasySep™小鼠CD4+CD25+调节性T细胞分选试剂盒II

免疫磁珠负选小鼠CD4+CD25+调节性T细胞分选试剂盒
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产品号 #（选择产品）

产品号 #18783_C

免疫磁珠负选小鼠CD4+CD25+调节性T细胞分选试剂盒

产品优势

  • 快速、易于操作
  • 纯度高达93%
  • 无需分离柱

产品组分包括

  • EasySep™小鼠CD4+ CD25+调节性T细胞分选试剂盒（产品号#18783）          
    • EasySep™小鼠CD4+ T细胞分选抗体混合物，0.5mL
    • EasySep™ Streptavidin RapidSpheres™ 50001磁珠，1mL          
    • EasySep™小鼠CD25调节性T细胞正选抗体混合物，0.5mL
    • EasySep™PE分选抗体混合物，1mL
    • EasySep™ Dextran RapidSpheres™ 50100 磁珠，1mL
    • EasySep™小鼠FcR阻断剂（产品号#18731），0.5mL          
  • RoboSep™小鼠CD4+ CD25+调节性T细胞分选试剂盒II（产品号#18783RF）          
    • EasySep™小鼠CD4+ T细胞分选抗体混合物，0.5mL
    • EasySep™ Streptavidin RapidSpheres™ 50001磁珠，1mL    
    • EasySep™小鼠CD25调节性T细胞正选抗体混合物，0.5mL
    • EasySep™PE分选抗体混合物，1mL
    • EasySep™ Dextran RapidSpheres™ 50100 磁珠，1mL
    • EasySep™小鼠FcR阻断剂（目录号#18731），0.5mL  
    • 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.
专为您的实验方案打造的产品
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. EasyEights™ EasySep™ Magnet
    EasyEights™ EasySep™ Magnet

    Multiple sample processing magnet for column-free immunomagnetic cell separation

  2. EasySep™ Buffer
    EasySep™ Buffer

    Cell separation buffer

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CD4+CD25+调节性T细胞分选试剂盒II，通过简单的两步法——先进行免疫磁珠负选，再进行正选，即可轻松从小鼠脾细胞或其他单细胞悬液样本中分离出高纯度的小鼠CD4+CD25+调节性T细胞（Tregs）。EasySep™技术结合单克隆抗体的特异性和无需分离柱的简便磁分选系统 ，已在发表的研究中广泛应用超过20年。

在此EasySep™细胞分选流程中，首先通过EasySep™小鼠CD4+T细胞分选抗体混合物进行负选 ，富集CD4+T细胞。随后使用EasySep™小鼠CD25调节性T细胞正选抗体 混合物，从预富集的细胞中正选出CD25+细胞。分选后的目的细胞 CD4+CD25+调节性T细胞即可用于流式细胞术、细胞培养、DNA/RNA提取等下游应用。

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、添加剂、抗体等。

在此EasySep™细胞分选流程中，首先通过EasySep™小鼠CD4+T细胞分选抗体混合物进行负选 ，富集CD4+T细胞。随后使用EasySep™小鼠CD25调节性T细胞正选抗体 混合物，从预富集的细胞中正选出CD25+细胞。分选后的目的细胞 CD4+CD25+调节性T细胞即可用于流式细胞术、细胞培养、DNA/RNA提取等下游应用。

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、添加剂、抗体等。

在此EasySep™细胞分选流程中，首先通过EasySep™小鼠CD4+T细胞分选抗体混合物进行负选 ，富集CD4+T细胞。随后使用EasySep™小鼠CD25调节性T细胞正选抗体 混合物，从预富集的细胞中正选出CD25+细胞。分选后的目的细胞 CD4+CD25+调节性T细胞即可用于流式细胞术、细胞培养、DNA/RNA提取等下游应用。

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、添加剂、抗体等。

在此EasySep™细胞分选流程中，首先通过EasySep™小鼠CD4+T细胞分选抗体混合物进行负选 ，富集CD4+T细胞。随后使用EasySep™小鼠CD25调节性T细胞正选抗体 混合物，从预富集的细胞中正选出CD25+细胞。分选后的目的细胞 CD4+CD25+调节性T细胞即可用于流式细胞术、细胞培养、DNA/RNA提取等下游应用。

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、添加剂、抗体等。

磁体兼容性
• EasySep™ Magnet (Catalog #18000) • “The Big Easy” EasySep™ Magnet (Catalog #18001) • EasyEights™ Magnet (Catalog #18103) • RoboSep™-S (Catalog #21000)
 
亚型
细胞分选试剂盒
 
细胞类型
T 细胞，T 细胞，调节性细胞
 
种属
小鼠
 
样本来源
Spleen
 
品牌
EasySep，RoboSep
 
研究领域
免疫
 

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实验数据

FACS Histogram Results with EasySep™ Mouse CD4+CD25+ Regulatory T Cell Isolation Kit II

Figure 1. Typical EasySep™ Mouse CD4+CD25+ Regulatory T Cell Isolation Profile

Starting with mouse splenocytes, the regulatory T cell content (CD4+CD25+FOXP3+) of the isolated fraction typically ranges from 70 - 93%. In the above example, the purities of the start and final isolated fractions are 2% and 72%, respectively.

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 1
Product Name
RoboSep™ Mouse CD4+CD25+ Regulatory T Cell Isolation Kit II
Catalog #
18783RF
Lot #
1000138480 or higher
Language
English
Document Type
Product Information Sheet 2
Product Name
RoboSep™ Mouse CD4+CD25+ Regulatory T Cell Isolation Kit II
Catalog #
18783RF
Lot #
1000138479 or lower
Language
English
Document Type
Product Information Sheet 1
Product Name
EasySep™ Mouse CD4+CD25+ Regulatory T Cell Isolation Kit II
Catalog #
18783
Lot #
1000138479 or lower
Language
English
Document Type
Product Information Sheet 2
Product Name
EasySep™ Mouse CD4+CD25+ Regulatory T Cell Isolation Kit II
Catalog #
18783
Lot #
1000138480 or higher
Language
English
Document Type
Safety Data Sheet 1
Product Name
RoboSep™ Mouse CD4+CD25+ Regulatory T Cell Isolation Kit II
Catalog #
18783RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Product Name
RoboSep™ Mouse CD4+CD25+ Regulatory T Cell Isolation Kit II
Catalog #
18783RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 3
Product Name
RoboSep™ Mouse CD4+CD25+ Regulatory T Cell Isolation Kit II
Catalog #
18783RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 4
Product Name
RoboSep™ Mouse CD4+CD25+ Regulatory T Cell Isolation Kit II
Catalog #
18783RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 5
Product Name
RoboSep™ Mouse CD4+CD25+ Regulatory T Cell Isolation Kit II
Catalog #
18783RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 6
Product Name
RoboSep™ Mouse CD4+CD25+ Regulatory T Cell Isolation Kit II
Catalog #
18783RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 7
Product Name
RoboSep™ Mouse CD4+CD25+ Regulatory T Cell Isolation Kit II
Catalog #
18783RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 8
Product Name
RoboSep™ Mouse CD4+CD25+ Regulatory T Cell Isolation Kit II
Catalog #
18783RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Product Name
EasySep™ Mouse CD4+CD25+ Regulatory T Cell Isolation Kit II
Catalog #
18783
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Product Name
EasySep™ Mouse CD4+CD25+ Regulatory T Cell Isolation Kit II
Catalog #
18783
Lot #
All
Language
English
Document Type
Safety Data Sheet 3
Product Name
EasySep™ Mouse CD4+CD25+ Regulatory T Cell Isolation Kit II
Catalog #
18783
Lot #
All
Language
English
Document Type
Safety Data Sheet 4
Product Name
EasySep™ Mouse CD4+CD25+ Regulatory T Cell Isolation Kit II
Catalog #
18783
Lot #
All
Language
English
Document Type
Safety Data Sheet 5
Product Name
EasySep™ Mouse CD4+CD25+ Regulatory T Cell Isolation Kit II
Catalog #
18783
Lot #
All
Language
English
Document Type
Safety Data Sheet 6
Product Name
EasySep™ Mouse CD4+CD25+ Regulatory T Cell Isolation Kit II
Catalog #
18783
Lot #
All
Language
English
Document Type
Safety Data Sheet 7
Product Name
EasySep™ Mouse CD4+CD25+ Regulatory T Cell Isolation Kit II
Catalog #
18783
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)

EasySep™ Cell Separation Technology
Brochure
EasySep™ Cell Separation Technology
Cell Enrichment Prior to Cell Sorting
Brochure
Cell Enrichment Prior to Cell Sorting
Tools for Your Immunology Research
Brochure
Tools for Your Immunology Research
Frequencies and Percentages of Mouse Immune Cell Types
Wallchart
Frequencies and Percentages of Mouse Immune Cell Types
Regulatory T Cells
Wallchart
Regulatory T Cells
Human Immune Cytokines
Wallchart
Human Immune Cytokines
How EasySep™ Magnetic Cell Separation Technology Works: Fast and Easy Cell Isolation
1:57
Video
How EasySep™ Magnetic Cell Separation Technology Works: Fast and Easy Cell Isolation
Isolate Cells with a Simple Pour-Off: EasySep™ Cell Separation Technology
1:13
Video
Isolate Cells with a Simple Pour-Off: EasySep™ Cell Separation Technology
Simultaneous Cell Isolation from Multiple Samples Using the EasyEights™ EasySep™ Magnet
0:57
Video
Simultaneous Cell Isolation from Multiple Samples Using the EasyEights™ EasySep™ Magnet
Streamline Your Mouse T Cell Research from Isolation to Expansion
30:18
Webinar
Streamline Your Mouse T Cell Research from Isolation to Expansion

常见问题

Can EasySep™ Streptavidin RapidSpheres™ be used for either positive or negative selection?

Currently, EasySep™ Streptavidin RapidSphere™ kits are only available for negative selection and work by targeting and removing unwanted cells.

How does the separation work?

Streptavidin RapidSphere™ magnetic particles are crosslinked to unwanted cells using biotinylated antibodies. When placed in the EasySep™ Magnet, labeled cells migrate to the wall of the tube. The unlabeled cells are then poured off into a new tube.

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™ Streptavidin RapidSphere™ separations be automated?

Yes. RoboSep™, the fully automated cell separator, automates all EasySep™ labeling and cell separation steps.

Are cells isolated using EasySep™ RapidSphere™ products FACS-compatible?

Yes. Desired cells are unlabeled and ready to use in downstream applications, such as FACS analysis.

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.

文献 (10)

Low-dose IL-2 prevents murine chronic cardiac allograft rejection: Role for IL-2-induced T regulatory cells and exosomes with PD-L1 and CD73. R. Ravichandran et al. American journal of transplantation : official journal of the American Society of Transplantation and the American Society of Transplant Surgeons 2022 sep

Abstract

To determine the effects and immunological mechanisms of low-dose interleukin-2 (IL-2) in a murine model of chronic cardiac allograft rejection (BALB/c to C57BL/6) after costimulatory blockade consisting of MR1 (250??$\mu$g/ip day 0) and CTLA4-Ig (200??$\mu$g/ip day 2), we administered low-dose IL-2 (2000??IU/day) starting on posttransplant day 14 for 3??weeks. T regulatory (Treg) cell infiltration of the grafts was determined by immunohistochemistry; circulating exosomes by western blot and aldehyde bead flow cytometry; antibodies to donor MHC by immunofluorescent staining of donor cells; and antibodies to cardiac self-antigens (myosin, vimentin) by ELISA. We demonstrated that costimulation blockade after allogeneic heart transplantation induced circulating exosomes containing cardiac self-antigens and antibodies to both donor MHC and self-antigens, leading to chronic rejection by day 45. Treatment with low-dose IL-2 prolonged allograft survival (>100??days), prevented chronic rejection, and induced splenic and graft-infiltrating CD4+ CD25+ Foxp3 Treg cells by day 45 and circulating exosomes (Foxp3+) with PD-L1 and CD73. MicroRNA 142, associated with the TGF$\beta$ pathway, was significantly downregulated in exosomes from IL-2-treated mice. In conclusion, low-dose IL-2 delays rejection in a murine model of chronic cardiac allograft rejection and also induces graft-infiltrating Tregs and circulating exosomes with immunoregulatory molecules.
View publication
Blockades of effector T cell senescence and exhaustion synergistically enhance antitumor immunity and immunotherapy. X. Liu et al. Journal for immunotherapy of cancer 2022 oct

Abstract

BACKGROUND Current immunotherapies still have limited successful rates among cancers. It is now recognized that T cell functional state in the tumor microenvironment (TME) is a key determinant for effective antitumor immunity and immunotherapy. In addition to exhaustion, cellular senescence in tumor-infiltrating T cells (TILs) has recently been identified as an important T cell dysfunctional state induced by various malignant tumors. Therefore, a better understanding of the molecular mechanism responsible for T cell senescence in the TME and development of novel strategies to prevent effector T cell senescence are urgently needed for cancer immunotherapy. METHODS Senescent T cell populations in the TMEs in mouse lung cancer, breast cancer, and melanoma tumor models were evaluated. Furthermore, T cell senescence induced by mouse tumor and regulatory T (Treg) cells in vitro was determined with multiple markers and assays, including real-time PCR, flow cytometry, and histochemistry staining. Loss-of-function strategies with pharmacological inhibitors and the knockout mouse model were used to identify the potential molecules and pathways involved in T cell senescence. In addition, melanoma mouse tumor immunotherapy models were performed to explore the synergistical efficacy of antitumor immunity via prevention of tumor-specific T cell senescence combined with anti-programmed death-ligand 1 (anti-PD-L1) checkpoint blockade therapy. RESULTS We report that both mouse malignant tumor cells and Treg cells can induce responder T cell senescence, similar as shown in human Treg and tumor cells. Accumulated senescent T cells also exist in the TME in tumor models of lung cancer, breast cancer and melanoma. Induction of ataxia-telangiectasia mutated protein (ATM)-associated DNA damage is the cause for T cell senescence induced by both mouse tumor cells and Treg cells, which is also regulated by mitogen-activated protein kinase (MAPK) signaling. Furthermore, blockages of ATM-associated DNA damage and/or MAPK signaling pathways in T cells can prevent T cell senescence mediated by tumor cells and Treg cells in vitro and enhance antitumor immunity and immunotherapy in vivo in adoptive transfer T cell therapy melanoma models. Importantly, prevention of tumor-specific T cell senescence via ATM and/or MAPK signaling inhibition combined with anti-PD-L1 checkpoint blockade can synergistically enhance antitumor immunity and immunotherapy in vivo. CONCLUSIONS These studies prove the novel concept that targeting both effector T cell senescence and exhaustion is an effective strategy and can synergistically enhance cancer immunotherapy.
View publication
Hypoxia-driven immunosuppression by Treg and type-2 conventional dendritic cells in HCC. S. Suthen et al. Hepatology (Baltimore, Md.) 2022 nov

Abstract

BACKGROUND AND AIMS Hypoxia is one of the central players in shaping the immune context of the tumor microenvironment (TME). However, the complex interplay between immune cell infiltrates within the hypoxic TME of HCC remains to be elucidated. APPROACH AND RESULTS We analyzed the immune landscapes of hypoxia-low and hypoxia-high tumor regions using cytometry by time of light, immunohistochemistry, and transcriptomic analyses. The mechanisms of immunosuppression in immune subsets of interest were further explored using in vitro hypoxia assays. Regulatory T cells (Tregs) and a number of immunosuppressive myeloid subsets, including M2 macrophages and human leukocyte antigen-DR isotype (HLA-DRlo ) type 2 conventional dendritic cell (cDC2), were found to be significantly enriched in hypoxia-high tumor regions. On the other hand, the abundance of active granzyme Bhi PD-1lo CD8+ T cells in hypoxia-low tumor regions implied a relatively active immune landscape compared with hypoxia-high regions. The up-regulation of cancer-associated genes in the tumor tissues and immunosuppressive genes in the tumor-infiltrating leukocytes supported a highly pro-tumorigenic network in hypoxic HCC. Chemokine genes such as CCL20 (C-C motif chemokine ligand 20) and CXCL5 (C-X-C motif chemokine ligand 5) were associated with recruitment of both Tregs and HLA-DRlo cDC2 to hypoxia-high microenvironments. The interaction between Tregs and cDC2 under a hypoxic TME resulted in a loss of antigen-presenting HLA-DR on cDC2. CONCLUSIONS We uncovered the unique immunosuppressive landscapes and identified key immune subsets enriched in hypoxic HCC. In particular, we identified a potential Treg-mediated immunosuppression through interaction with a cDC2 subset in HCC that could be exploited for immunotherapies.
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更多信息

更多信息
种属 Mouse
Magnet Compatibility • EasySep™ Magnet (Catalog #18000) • “The Big Easy” EasySep™ Magnet (Catalog #18001) • EasyEights™ Magnet (Catalog #18103) • RoboSep™-S (Catalog #21000)
样本来源 Spleen
标记抗体
  1. 抗小鼠CD4抗体，克隆RM4-5
    抗小鼠CD4抗体，克隆RM4-5

    大鼠单克隆IgG2a抗体，抗小鼠CD4

  2. 抗小鼠CD4抗体，克隆RM4-4
    抗小鼠CD4抗体，克隆RM4-4

    抗小鼠CD4的大鼠单克隆IgG2b抗体

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