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。

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。

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。

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。

磁体兼容性
• 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)

亚型
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

筛选方法
Negative

应用
细胞分选

品牌
EasySep，RoboSep

研究领域
免疫

查看更多显示更少

实验数据

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

Workflow Stages for T Cell Research

Cell Sourcing

Cell Isolation

Cell Activation, Expansion, and Differentiation

Cell Characterization

Workflow Stages for T Cell Engineering

Cell Isolation

Cell Activation, Expansion, and Differentiation

Cell Characterization

Cell Cryopreservation

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.

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.

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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."

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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.

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EasySep™人记忆CD4+ T细胞富集试剂盒

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产品号 #19157_C

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Scientists Helping Scientists™

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

EasySep™人记忆CD4+ T细胞富集试剂盒

产品号 #（选择产品）

产品号 #19157_C

产品优势

产品组分包括

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

技术资料 (10)

常见问题

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

How does the separation work?

Which columns do I use?

How can I analyze the purity of my enriched sample?

Can EasySep™ separations be automated?

Can EasySep™ be used to isolate rare cells?

Are the EasySep™ magnetic particles FACS-compatible?

Can the EasySep™ magnetic particles be removed after enrichment?

Can I alter the separation time in the magnet?

For positive selection, can I perform more than 3 separations to increase purity?

How does the binding of the EasySep™ magnetic particle affect the cells? is the function of positively selected cells altered by the bound particles?

文献 (5)

Abstract

Abstract

Abstract

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