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EasySep™人记忆CD4+ T细胞富集试剂盒

未标记的人记忆 CD4+ T细胞的免疫磁珠负选
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产品号 #（选择产品）

产品号 #19157_C

未标记的人记忆 CD4+ T细胞的免疫磁珠负选

产品优势

  • 操作简单、快捷，且无需分离柱
  • 纯度高达98%
  • 获得的活细胞无标记

产品组分包括

  • EasySep™  人记忆CD4+ T细胞富集试剂盒（产品号 #19157）
    • EasySep™人记忆CD4+ T细胞富集抗体混合物，1 mL
    • EasySep™ D Magnetic Particles磁珠，2 x 1 mL
  • RoboSep™ 人 记忆CD4+ T细胞富集试剂盒（含滤芯吸头）（产品号 #19157RF）
    • EasySep™人记忆CD4+ T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）
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. EasySep™磁铁
    EasySep™磁铁
  2. EasySep™缓冲液
    EasySep™缓冲液
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™人记忆CD4+ T细胞富集试剂盒，通过免疫磁珠负选，可轻松高效地从新鲜或冻存的人外周血单个核细胞（PBMC）样本中分离高纯度的人记忆 CD4+ T细胞（CD4+CD45RA-CD45RO+）。EasySep™结合了单克隆抗体的特异性和无柱磁性系统的简便性，迄今已广泛应用于发表的研究中超过20年。    

在此 EasySep™负选过程中，表达以下标记物的非目的细胞通过抗体复合物与磁珠标记 被靶向去除：CD8、CD14、CD16、CD19、CD20、CD36、CD45RA、CD56、CD123、TCRγ/δ及glyA。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管中即可。经磁珠分选获得的记忆CD4+ T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。

了解更多关于免疫磁珠EasySep™技术的工作原理，或如何通过RoboSep™实现全自动化免疫磁珠细胞分选 。或选择经EasySep™人记忆CD4+ T细胞富集试剂盒分离的、符合伦理规范 的即用型冻存人外周血CD4+CD45RO+ T细胞 。探索更多优化您实验流程的产品，包括培养基、添加剂、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） • EasyPlate™ EasySep™磁极（产品号 #18102） • Easy 50 EasySep™磁极（产品号 #18002） • RoboSep™-S（产品号 #21000）
 
分类
细胞分选试剂盒
 
细胞类型
T 细胞，T 细胞，CD4+
 
种属
人
 
样本来源
PBMC
 
分选方法
负选
 
应用
细胞分选
 
品牌
EasySep，RoboSep
 
研究领域
免疫
 

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实验数据

Typical Enrichment Profile For EasySep™ Human Memory CD4+ T Cell Enrichment Kit

Figure 1. Typical Enrichment Profile For EasySep™ Human Memory CD4+ T Cell Enrichment Kit

Starting with previously frozen mononuclear cells, the memory CD4+ T cell content of the enriched fraction typically ranges from 86% - 98%.

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
Product Name
RoboSep™ Human Memory CD4 T Cell Enrichment Kit with Filter Tips
Catalog #
19157RF
Lot #
All
Language
English
Document Type
Product Information Sheet
Product Name
EasySep™ Human Memory CD4+ T Cell Enrichment Kit
Catalog #
19157
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Product Name
RoboSep™ Human Memory CD4 T Cell Enrichment Kit with Filter Tips
Catalog #
19157RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Product Name
RoboSep™ Human Memory CD4 T Cell Enrichment Kit with Filter Tips
Catalog #
19157RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 3
Product Name
RoboSep™ Human Memory CD4 T Cell Enrichment Kit with Filter Tips
Catalog #
19157RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Product Name
EasySep™ Human Memory CD4+ T Cell Enrichment Kit
Catalog #
19157
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Product Name
EasySep™ Human Memory CD4+ T Cell Enrichment Kit
Catalog #
19157
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)

Tools for Your Immunology Research
Brochure
Tools for Your Immunology Research
Human T Cell Research Product Workflow
Brochure
Human T Cell Research Product Workflow
EasySep™ Cell Separation Technology
Brochure
EasySep™ Cell Separation Technology
Antigen Processing and Presentation
Wallchart
Antigen Processing and Presentation
Human Immune Cytokines
Wallchart
Human Immune Cytokines
Frequencies of Human Cell Types in Blood-Related Sources
Wallchart
Frequencies of Human Cell Types in Blood-Related Sources
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
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
How to Isolate PBMCs from Whole Blood Using Density Gradient Centrifugation (Ficoll™ or Lymphoprep™)
1:37
Video
How to Isolate PBMCs from Whole Blood Using Density Gradient Centrifugation (Ficoll™ or Lymphoprep...
Cell Isolation of Human CD4+ Memory T Cells By Column-Free Immunomagnetic Cell Separation
Scientific Poster
Cell Isolation of Human CD4+ Memory T Cells By Column-Free Immunomagnetic Cell Separation

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 (5)

The human liver microenvironment shapes the homing and function of CD4+ T-cell populations. B. G. Wiggins et al. Gut 2022 jul

Abstract

OBJECTIVE Tissue-resident memory T cells (TRM) are vital immune sentinels that provide protective immunity. While hepatic CD8+ TRM have been well described, little is known about the location, phenotype and function of CD4+ TRM. DESIGN We used multiparametric flow cytometry, histological assessment and novel human tissue coculture systems to interrogate the ex vivo phenotype, function and generation of the intrahepatic CD4+ T-cell compartment. We also used leukocytes isolated from human leukocyte antigen (HLA)-disparate liver allografts to assess long-term retention. RESULTS Hepatic CD4+ T cells were delineated into three distinct populations based on CD69 expression: CD69-, CD69INT and CD69HI. CD69HICD4+ cells were identified as tissue-resident CD4+ T cells on the basis of their exclusion from the circulation, phenotypical profile (CXCR6+CD49a+S1PR1-PD-1+) and long-term persistence within the pool of donor-derived leukcoocytes in HLA-disparate liver allografts. CD69HICD4+ T cells produced robust type 1 polyfunctional cytokine responses on stimulation. Conversely, CD69INTCD4+ T cells represented a more heterogenous population containing cells with a more activated phenotype, a distinct chemokine receptor profile (CX3CR1+CXCR3+CXCR1+) and a bias towards interleukin-4 production. While CD69INTCD4+ T cells could be found in the circulation and lymph nodes, these cells also formed part of the long-term resident pool, persisting in HLA-mismatched allografts. Notably, frequencies of CD69INTCD4+ T cells correlated with necroinflammatory scores in chronic hepatitis B infection. Finally, we demonstrated that interaction with hepatic epithelia was sufficient to generate CD69INTCD4+ T cells, while additional signals from the liver microenvironment were required to generate liver-resident CD69HICD4+ T cells. CONCLUSIONS High and intermediate CD69 expressions mark human hepatic CD4+ TRM and a novel functionally distinct recirculating population, respectively, both shaped by the liver microenvironment to achieve diverse immunosurveillance.
View publication
Stress hormone signalling inhibits Th1 polarization in a CD4 T-cell-intrinsic manner via mTORC1 and the circadian gene PER1. C. M. Capelle et al. Immunology 2022 apr

Abstract

Stress hormones are believed to skew the CD4 T-cell differentiation towards a Th2 response via a T-cell-extrinsic mechanism. Using isolated primary human na{\{i}}ve and memory CD4 T cells here we show that both adrenergic- and glucocorticoid-mediated stress signalling pathways play a CD4 na{\"{i}}ve T-cell-intrinsic role in regulating the Th1/Th2 differentiation balance. Both stress hormones reduced the Th1 programme and cytokine production by inhibiting mTORC1 signalling via two parallel mechanisms. Stress hormone signalling inhibited mTORC1 in na{\"{i}}ve CD4 T cells (1) by affecting the PI3K/AKT pathway and (2) by regulating the expression of the circadian rhythm gene period circadian regulator 1 (PER1). Both stress hormones induced the expression of PER1 which inhibited mTORC1 signalling thus reducing Th1 differentiation. This previously unrecognized cell-autonomous mechanism connects stress hormone signalling with CD4 T-cell differentiation via mTORC1 and a specific circadian clock gene namely PER1."
View publication
PD-1 blockade potentiates HIV latency reversal ex vivo in CD4+ T cells from ART-suppressed individuals. R. Fromentin et al. Nature communications 2019 feb

Abstract

HIV persists in latently infected CD4+ T cells during antiretroviral therapy (ART). Immune checkpoint molecules, including PD-1, are preferentially expressed at the surface of persistently infected cells. However, whether PD-1 plays a functional role in HIV latency and reservoir persistence remains unknown. Using CD4+ T cells from HIV-infected individuals, we show that the engagement of PD-1 inhibits viral production at the transcriptional level and abrogates T-cell receptor (TCR)-induced HIV reactivation in latently infected cells. Conversely, PD-1 blockade with the monoclonal antibody pembrolizumab enhances HIV production in combination with the latency reversing agent bryostatin without increasing T cell activation. Our results suggest that the administration of immune checkpoint blockers to HIV-infected individuals on ART may facilitate latency disruption.
View publication
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更多信息

更多信息
种属 Human
Magnet Compatibility • EasySep™ Magnet (Catalog #18000) • “The Big Easy” EasySep™ Magnet (Catalog #18001) • EasyPlate™ EasySep™ Magnet (Catalog 18102) • Easy 50 EasySep™ Magnet (Catalog #18002) • RoboSep™-S (Catalog #21000)
样本来源 PBMC
Selection Method Negative
标记抗体
  1. 抗人CD45RO抗体，克隆UCHL1
    抗人CD45RO抗体，克隆UCHL1

    小鼠单克隆IgG2a抗体，抗人、黑猩猩、普通狨猴CD45RO

  2. 抗人CD4抗体，克隆OKT4
    抗人CD4抗体，克隆OKT4

    小鼠单克隆IgG2b抗体，抗人、恒河猴、食蟹猴CD4

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