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EasySep™人记忆CD4+ T细胞富集试剂盒

未触及人记忆CD4+ T细胞的免疫磁阴性分离
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产品号 #19157_C

未触及人记忆CD4+ T细胞的免疫磁阴性分离

产品优势

  • 快速，易于使用和无列
  • 纯度高达98%
  • 未接触的活细胞

产品组分包括

  • EasySep™人记忆CD4+ T细胞富集试剂盒（目录#19157）
    • EasySep™人记忆CD4+ T细胞浓缩鸡尾酒，1ml
    • EasySep™D磁性颗粒，2 × 1ml
  • RoboSep™人类记忆CD4 T细胞富集试剂盒与过滤器提示（目录#19157RF）
    • EasySep™人记忆CD4+ T细胞浓缩鸡尾酒，1ml
    • EasySep™D磁性颗粒，2 × 1ml
    • RoboSep™缓冲器（目录#20104）
    • RoboSep™过滤器提示(目录#20125)
main product photo
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Products for Your Protocol
To see all required products for your protocol, please consult the Protocols and Documentation.
  1. EasySep™ Magnet
    EasySep™ Magnet

    Magnet for column-free immunomagnetic 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

概述

使用EasySep™人记忆CD4+ T细胞富集试剂盒，通过免疫磁阴性选择，从新鲜或先前冷冻的人外周血单个核细胞（PBMC）样品中轻松高效地分离高度纯化的人记忆CD4+ T细胞（CD4+CD45RA-CD45RO+）。EasySep™在已发表的研究中广泛使用了20多年，它结合了单克隆抗体的特异性和无柱磁系统的简单性。

在这个EasySep™阴性选择程序中，用抗体复合物和磁性颗粒标记不需要的细胞。表达以下标记的不需要的细胞被靶向去除：CD8， CD14, CD16, CD19, CD20, CD36, CD45RA, CD56, CD123， TCRγ/δ和glyA。然后使用EasySep™磁铁将磁性标记的细胞与未触碰的所需记忆CD4+ T细胞分离，并简单地将所需细胞倒入或移液到新的管中。磁性细胞分离后，所需的记忆CD4+ T细胞可用于下游应用，例如流式细胞术，培养或DNA/RNA提取。

了解更多关于免疫磁性的知识EasySep™技术工作原理或如何完全自动化免疫磁细胞分离RoboSep™． 或者，选择现成的、道德来源的、主要的人外周血CD4+CD45RO+ T细胞，冷冻使用EasySep™Human Memory CD4+ T细胞富集试剂盒进行分离。探索额外的产品针对您的工作流程进行了优化，包括培养基、补充剂、抗体等。

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)
 
Subtype
Cell Isolation Kits
 
Cell Type
T Cells, T Cells, CD4+
 
Species
Human
 
Sample Source
PBMC
 
Selection Method
Negative
 
Application
Cell Isolation
 
Brand
EasySep, RoboSep
 
Area of Interest
Immunology
 
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Data Figures

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%.

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
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

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 (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

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 (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
Species 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)
Sample Source 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

    小鼠抗人、黑猩猩、普通狨猴CD45RO单克隆IgG2a抗体

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

    小鼠抗人、恒河猴、食蟹CD4单克隆IgG2b抗体

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