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EasySep™人CD4正选试剂盒II

人CD4+细胞的免疫磁珠正选
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产品号 #（选择产品）

产品号 #17852_C

人CD4+细胞的免疫磁珠正选

产品优势

  • 快捷、操作简单
  • 纯度高达96%
  • 无需分离柱

产品组分包括

  • EasySep™人CD4正选试剂盒II（产品号 ＃17852）
    • EasySep™人CD4正选抗体混合物II，0.5mL
    • EasySep™ Dextran RapidSpheres™ 50100 磁珠,1mL
  • EasySep™人CD4正选试剂盒II（产品号 #100-0693）
    • EasySep™人CD4正选抗体混合物II，1x5mL
    • EasySep™ Dextran RapidSpheres™ 50100 磁珠，2x1mL
  • RoboSep™人CD4正选试剂盒II（产品号 ＃17852RF）
    • EasySep™人CD4正选抗体混合物II，0.5mL
    • EasySep™ Dextran RapidSpheres™ 50100 磁珠，1mL
    • RoboSep™ 缓冲液（产品号 #20104）
    • RoboSep™过滤吸头（目录编号20125）
main product photo
New look, same high quality and support! You may notice that your instrument or reagent packaging looks slightly different from images displayed on the website, or from previous orders. We are updating our look but rest assured, the products themselves and how you should use them have not changed. Learn more
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
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总览

通过免疫磁珠正选，EasySep™人CD4正选试剂盒II可以从新鲜或冻存的人外周血单个核细胞 (PBMCs) 或洗涤的白细胞单采术样本中分离出高度纯化的 CD4+ 细胞。EasySep™技术结合单克隆抗体的特异性和免磁柱系统的简便性，已在发表的研究中被广泛应用超过20年。

在本款EasySep™阳性分选流程中，目标细胞通过与识别CD4的抗体复合物及磁珠进行标记。使用EasySep™磁力架分离标记细胞，只需倾倒或枪头吸取非目标细胞。目标细胞保留在管中。仅需短至15分钟磁珠分选后，目标CD4+细胞即可直接用于流式细胞术、培养或DNA/RNA提取等下游应用。

该产品可替代EasySep™人CD4正选试剂盒 (产品号 #18052) 以进行更快的细胞分选。

如需从大量白细胞分离样本中分选CD4+细胞，请选用大容量规格（1x10^10细胞）试剂盒（产品号 #100-0693）。

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) • Easy 50 EasySep™ Magnet (Catalog #18002) • EasyEights™ EasySep™ Magnet (Catalog #18103) • RoboSep™-S (Catalog #21000) • Easy 250 EasySep™ Magnet (Catalog #100-0821)
 
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
Leukapheresis，PBMC
 
筛选方法
Positive
 
应用
细胞分选
 
品牌
EasySep，RoboSep
 
研究领域
免疫
 

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Data Figures

Typical EasySep™ Human CD4 Positive Selection Profile

Figure 1. Typical EasySep™ Human CD4 Positive Selection Profile

Starting with human PBMCs, the CD4+ T cell content of the isolated fraction is typically 90 ± 6% (mean ± SD) using the purple EasySep™ Magnet.

FACS Data for Anti-Human CD4 Antibody, Clone OKT4, Alexa Fluor® 488-Conjugated

Figure 2. FACS Data for Anti-Human CD4 Antibody, Clone OKT4, Alexa Fluor® 488-Conjugated

(A) Flow cytometry analysis of human peripheral blood mononuclear cells (PBMCs) labeled with Anti-Human CD4 Antibody, Clone OKT4, Alexa Fluor® 488 (Catalog #60016AD) and Anti-Human CD45 Antibody, Clone HI30, APC (Catalog #60018AZ).

(B) Flow cytometry analysis of human PBMCs isolated with the EasySep™ Human CD3 Positive Selection Kit (Catalog #17851) and labeled with Anti-Human CD4 Antibody, Clone OKT4, Alexa Fluor® 488. Histograms show labeling of total PBMCs (Start) and isolated cells (Isolated). Labeling with Mouse IgG2b, kappa Isotype Control Antibody, Clone MPC-11, Alexa Fluor® 488 (Catalog #60072AD) is shown in the bottom panel (solid line histogram).

(C) Flow cytometry analysis of human PBMCs isolated with the EasySep™ Human CD4 Positive Selection Kit (Catalog #17852) and labeled with Anti-Human CD4 Antibody, Clone OKT4, Alexa Fluor® 488. Histograms show labeling of total PBMCs (Start) and isolated cells (Isolated). Labeling with Mouse IgG2b, kappa Isotype Control Antibody, Clone MPC-11, Alexa Fluor® 488 is shown in the bottom panel (solid line histogram).

Gene Expression Profiles of EasySep™-Isolated CD4+ T Cells Are Similar to PBMC Control

Figure 3. Gene Expression Profiles of EasySep™-Isolated CD4+ T Cells Are Similar to PBMC Control

(A,B) tSNE plots were generated using data from (A) PBMC control or (B) cells isolated using the EasySep™ Human CD4+ T Cell Enrichment Kit (Catalog #19052). CD4+ T cell clusters are colored as indicated in the legend.

(C,D) 500 genes were selected from a previously published list of CD4+ T cell signature markers (Zhang et al., 2018). Expression heatmaps were generated for CD4+ cells from (C) PBMC control and (D) cells isolated using the EasySep™ Human CD4 Positive Selection Kit II (Catalog #17852), EasySep™ Human CD4+ T Cell Enrichment Kit (Catalog #19052), or the EasySep™ Release Human CD4 Positive Selection Kit (Catalog #17752). The average expression was calculated within each sample for three CD4+ T cell clusters identified by Seurat (naïve, central memory, and effector memory CD4 T+ cells).

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
EasySep™ Human CD4 Positive Selection Kit II
Catalog #
17852
Lot #
All
Language
English
Document Type
Product Information Sheet
Product Name
RoboSep™ Human CD4 Positive Selection Kit II
Catalog #
17852RF
Lot #
All
Language
English
Document Type
Product Information Sheet
Product Name
EasySep™ Human CD4 Positive Selection Kit II
Catalog #
100-0693
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Product Name
EasySep™ Human CD4 Positive Selection Kit II
Catalog #
17852
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Product Name
EasySep™ Human CD4 Positive Selection Kit II
Catalog #
17852
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Product Name
RoboSep™ Human CD4 Positive Selection Kit II
Catalog #
17852RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Product Name
RoboSep™ Human CD4 Positive Selection Kit II
Catalog #
17852RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 3
Product Name
RoboSep™ Human CD4 Positive Selection Kit II
Catalog #
17852RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Product Name
EasySep™ Human CD4 Positive Selection Kit II
Catalog #
100-0693
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Product Name
EasySep™ Human CD4 Positive Selection Kit II
Catalog #
100-0693
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)

EasySep™ Cell Separation Technology
Brochure
EasySep™ Cell Separation Technology
Isolate Human Immune Cells
Brochure
Isolate Human Immune Cells
Tools for Your Immunology Research
Brochure
Tools for Your Immunology Research
Frequencies of Human Cell Types in Blood-Related Sources
Wallchart
Frequencies of Human Cell Types in Blood-Related Sources
The Immune Response to HIV Poster
Wallchart
The Immune Response to HIV Poster
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
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...

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 (6)

Engagement of TRAIL triggers degranulation and IFN$\gamma$ production in human natural killer cells. J. H\ofle et al." EMBO reports 2022 aug

Abstract

NK cells utilize a large array of receptors to screen their surroundings for aberrant or virus-infected cells. Given the vast diversity of receptors expressed on NK cells we seek to identify receptors involved in the recognition of HIV-1-infected cells. By combining an unbiased large-scale screening approach with a functional assay, we identify TRAIL to be associated with NK cell degranulation against HIV-1-infected target cells. Further investigating the underlying mechanisms, we demonstrate that TRAIL is able to elicit multiple effector functions in human NK cells independent of receptor-mediated induction of apoptosis. Direct engagement of TRAIL not only results in degranulation but also IFN$\gamma$ production. Moreover, TRAIL-mediated NK cell activation is not limited to its cognate death receptors but also decoy receptor I, adding a new perspective to the perceived regulatory role of decoy receptors in TRAIL-mediated cytotoxicity. Based on these findings, we propose that TRAIL not only contributes to the anti-HIV-1 activity of NK cells but also possesses a multifunctional role beyond receptor-mediated induction of apoptosis, acting as a regulator for the induction of different effector functions.
View publication
Targeting 3D chromosomal architecture at the RANK loci to suppress myeloma-driven osteoclastogenesis. K. Th\ummler et al." Oncoimmunology 2022

Abstract

Bone disease represents a major cause of morbidity and mortality in Multiple Myeloma (MM); primarily driven by osteoclasts whose differentiation is dependent on expression of RANKL by MM cells. Notably, costimulation by ITAM containing receptors (i.e., Fc$\gamma$R) can also play a crucial role in osteoclast differentiation. Modeling the pathology of the bone marrow microenvironment with an ex vivo culture system of primary human multiple myeloma cells, we herein demonstrate that Fc$\gamma$R-mediated signaling, via staphylococcal protein A (SpA) IgG immune-complexes, can act as a critical negative regulator of MM-driven osteoclast differentiation. Interrogation of the mode-of-action revealed that Fc$\gamma$R-mediated signaling causes epigenetic modulation of chromosomal 3D architecture at the RANK promoter; with altered spatial orientation of a proximal super enhancer. Combined this leads to substantial down-regulation of RANK at a transcript, protein, and functional level. These observations shed light on a novel mechanism regulating RANK expression and provide a rationale for targeting Fc$\gamma$R-signaling for the amelioration of osteolytic bone pathology in disease.
View publication
Competition between PAF1 and MLL1/COMPASS confers the opposing function of LEDGF/p75 in HIV latency and proviral reactivation. R. Gao et al. Science advances 2020 may

Abstract

Transcriptional status determines the HIV replicative state in infected patients. However, the transcriptional mechanisms for proviral replication control remain unclear. In this study, we show that, apart from its function in HIV integration, LEDGF/p75 differentially regulates HIV transcription in latency and proviral reactivation. During latency, LEDGF/p75 suppresses proviral transcription via promoter-proximal pausing of RNA polymerase II (Pol II) by recruiting PAF1 complex to the provirus. Following latency reversal, MLL1 complex competitively displaces PAF1 from the provirus through casein kinase II (CKII)-dependent association with LEDGF/p75. Depleting or pharmacologically inhibiting CKII prevents PAF1 dissociation and abrogates the recruitment of both MLL1 and Super Elongation Complex (SEC) to the provirus, thereby impairing transcriptional reactivation for latency reversal. These findings, therefore, provide a mechanistic understanding of how LEDGF/p75 coordinates its distinct regulatory functions at different stages of the post-integrated HIV life cycles. Targeting these mechanisms may have a therapeutic potential to eradicate HIV infection.
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) • Easy 50 EasySep™ Magnet (Catalog #18002) • EasyEights™ EasySep™ Magnet (Catalog #18103) • RoboSep™-S (Catalog #21000) • Easy 250 EasySep™ Magnet (Catalog #100-0821)
Sample Source Leukapheresis, PBMC
Selection Method Positive
标记抗体
  1. 抗人CD8a抗体，克隆RPA-T8
    抗人CD8a抗体，克隆RPA-T8

    小鼠单克隆IgG1抗体，抗人、恒河猴、食蟹猴CD8a

  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

  3. 山羊抗小鼠 IgG (H+L) 抗体，多克隆
    山羊抗小鼠 IgG (H+L) 抗体，多克隆

    抗小鼠IgG（H+L）山羊多克隆抗体

  4. 抗人CD3抗体，克隆UCHT1
    抗人CD3抗体，克隆UCHT1

    小鼠（BALB/c）单克隆IgG1抗体，抗人、黑猩猩CD3

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  • ISO 13485 和 ISO 9001 质量管理体系认证
    ISO 14001 环境管理体系认证

Scientists Helping Scientists™

ISO 13485 和 ISO 9001 质量管理体系认证

ISO 14001 环境管理体系认证

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