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EasySep™人CD4+ T细胞分选试剂盒

免疫磁珠负选人CD4+ T细胞
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产品号 #（选择产品）

产品号 #17952_C

免疫磁珠负选人CD4+ T细胞

产品优势

  • 操作简单、快捷，且无需分离柱
  • 纯度高达97%，回收率高
  • 获得不带标记的活细胞    

产品组分包括

  • EasySep™人CD4+ T细胞分选试剂盒（产品号 #17952）
    • EasySep™人CD4+ T细胞分选抗体混合物, 1 mL
    • EasySep™ Dextran RapidSpheres™磁珠, 1 mL
  • EasySep™人CD4+ T细胞分选试剂盒（产品号 #100-0696）
    • EasySep™人CD4+ T细胞分选抗体混合物, 1 x 10 mL
    • EasySep™ Dextran RapidSpheres™磁珠, 1 x 10 mL
  • RoboSep™人CD4+ T细胞分选试剂盒（产品号 #17952RF）
    • EasySep™人CD4+ T细胞分选抗体混合物, 1 mL
    • EasySep™ Dextran RapidSpheres™磁珠, 1 mL
    • RoboSep™ 缓冲液（产品号 #20104）
    • RoboSep™过滤吸头（产品号 #20125）
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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™磁极
    EasySep™磁极
  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CD4+ T细胞分选试剂盒，可轻松高效地从新鲜或冻存的人外周血单个核细胞（PBMCs）或白细胞单采样本中通过免疫磁珠负选获得高纯度人CD4+ T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，已在发表的研究中广泛应用超过20年。

在该 EasySep™负选流程中，非目的细胞会被抗体复合物与磁珠标记。表达以下标记物的非目的细胞将被靶向去除：CD8、CD14、CD16、CD19、CD36、CD56、CD66b、CD123、TCRgd 和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，将目的细胞简单倾倒或移液吸取至新试管中。仅需8分钟即可完成磁珠分选，获得的 CD4+ T细胞可用于流式细胞术、细胞培养或DNA/RNA提取等下游应用。

该试剂盒可替代EasySep™人CD4+ T细胞富集试剂盒 (产品号 #19052) 以实现更快速地细胞分选。

如需从白细胞单采样本中大规模分选人CD4+ T细胞，请选用大包装（1×10^10细胞）试剂盒（产品号 #100-0696）。

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。或直接选用 EasySep™人CD4+ 分选试剂盒提供的即用型、符合伦理道德的冻存人外周血CD4+ T原代细胞。探索其他针对您的工作流程优化的产品，包括培养基、补充剂、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）
• EasyEights™ EasySep™磁极（产品号 #18103）
• RoboSep™-S（产品号 #21000）
• Easy 250 EasySep™磁极（产品号 #100-0821）
 
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
白细胞单采术样本、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

CD4+ T cell separation using EasySep™ Human CD4+ T Cell Isolation Kit

Figure 1. EasySep™ Human CD4+ T Cell Isolation Kit

Starting with human peripheral blood mononuclear cells (PBMCs), the CD4+ T cell (CD3+CD4+) content of the isolated fraction is typically 94.8 ± 2.3% (mean ± SD).

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
EasySep™ Human CD4+ T Cell Isolation Kit
Catalog #
100-0696
Lot #
All
Language
English
Document Type
Product Information Sheet 1
Product Name
EasySep™ Human CD4+ T Cell Isolation Kit
Catalog #
17952
Lot #
All
Language
Multi
Document Type
Product Information Sheet 2
Product Name
EasySep™ Human CD4+ T Cell Isolation Kit
Catalog #
17952
Lot #
All
Language
English
Document Type
Product Information Sheet 1
Product Name
RoboSep™ Human CD4+ T Cell Isolation Kit
Catalog #
17952RF
Lot #
All
Language
English
Document Type
Product Information Sheet 2
Product Name
RoboSep™ Human CD4+ T Cell Isolation Kit
Catalog #
17952RF
Lot #
All
Language
Multi
Document Type
Safety Data Sheet 1
Product Name
EasySep™ Human CD4+ T Cell Isolation Kit
Catalog #
17952
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Product Name
EasySep™ Human CD4+ T Cell Isolation Kit
Catalog #
17952
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Product Name
RoboSep™ Human CD4+ T Cell Isolation Kit
Catalog #
17952RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Product Name
RoboSep™ Human CD4+ T Cell Isolation Kit
Catalog #
17952RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 3
Product Name
RoboSep™ Human CD4+ T Cell Isolation Kit
Catalog #
17952RF
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 (15)

Tools for Your Immunology Research
Brochure
Tools for Your Immunology Research
EasySep™ Cell Separation Technology
Brochure
EasySep™ Cell Separation Technology
Isolate Human Immune Cells
Brochure
Isolate Human Immune Cells
Cell Enrichment Prior to Cell Sorting
Brochure
Cell Enrichment Prior to Cell Sorting
The Immune Response to HIV Poster
Wallchart
The Immune Response to HIV Poster
Production of Chimeric Antigen Receptor T Cells
Wallchart
Production of Chimeric Antigen Receptor T Cells
Antigen Processing and Presentation
Wallchart
Antigen Processing and Presentation
Human Immune Cytokines
Wallchart
Human Immune Cytokines
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 to Isolate Cells with EasySep™ Column-Free Cell Separation Technology
1:23
Video
How to Isolate Cells with EasySep™ Column-Free Cell Separation Technology
How to Isolate Cells in 96-Well Plates Using the EasyPlate™ Cell Separation Magnet
2:34
Video
How to Isolate Cells in 96-Well Plates Using the EasyPlate™ Cell Separation Magnet
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
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...
Next-Generation Tools to Advance Your Human T Cell Research
32:50
Webinar
Next-Generation Tools to Advance Your Human T Cell Research

常见问题 (11)

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 (46)

Release of P-TEFb from the Super Elongation Complex promotes HIV-1 latency reversal PLOS Pathogens 2024 Sep

Abstract

The persistence of HIV-1 in long-lived latent reservoirs during suppressive antiretroviral therapy (ART) remains one of the principal barriers to a functional cure. Blocks to transcriptional elongation play a central role in maintaining the latent state, and several latency reversal strategies focus on the release of positive transcription elongation factor b (P-TEFb) from sequestration by negative regulatory complexes, such as the 7SK complex and BRD4. Another major cellular reservoir of P-TEFb is in Super Elongation Complexes (SECs), which play broad regulatory roles in host gene expression. Still, it is unknown if the release of P-TEFb from SECs is a viable latency reversal strategy. Here, we demonstrate that the SEC is not required for HIV-1 replication in primary CD4+ T cells and that a small molecular inhibitor of the P-TEFb/SEC interaction (termed KL-2) increases viral transcription. KL-2 acts synergistically with other latency reversing agents (LRAs) to reactivate viral transcription in several cell line models of latency in a manner that is, at least in part, dependent on the viral Tat protein. Finally, we demonstrate that KL-2 enhances viral reactivation in peripheral blood mononuclear cells (PBMCs) from people living with HIV (PLWH) on suppressive ART, most notably in combination with inhibitor of apoptosis protein antagonists (IAPi). Taken together, these results suggest that the release of P-TEFb from cellular SECs may be a novel route for HIV-1 latency reactivation. Author summarySince the start of the HIV pandemic, it is estimated that nearly 86 million people have been infected with the virus, and about 40 million people have died. Modern antiretroviral therapies potently restrict viral replication and prevent the onset of AIDS, saving millions of lives. However, these therapies are not curative due to the persistence of the virus in a silenced or ‘latent’ state in long-lived cells of the body. One proposed strategy to clear this latent reservoir, termed “shock and kill”, is to activate these silenced viruses such that the infected cells can be cleared from the body by the immune system. While several drugs have been developed that can activate latent viruses, none have proven effective at reducing the size of the latent reservoir in patients in clinical trials. Here, we describe a new method for latency reactivation using a small molecule inhibitor of a human protein complex called the Super Elongation Complex (SEC). Inhibiting the SEC enhances viral transcription during active infection and triggers the reactivation of latent viruses, especially when in combination with other latency reversing agents. These results pave the way for developing more effective strategies to reactivate latent viruses towards a functional cure.
View publication
Integrated Single-cell Multiomic Analysis of HIV Latency Reversal Reveals Novel Regulators of Viral Reactivation Genomics, Proteomics & Bioinformatics 2024 Jun

Abstract

AbstractDespite the success of antiretroviral therapy, human immunodeficiency virus (HIV) cannot be cured because of a reservoir of latently infected cells that evades therapy. To understand the mechanisms of HIV latency, we employed an integrated single-cell RNA sequencing (scRNA-seq) and single-cell assay for transposase-accessible chromatin with sequencing (scATAC-seq) approach to simultaneously profile the transcriptomic and epigenomic characteristics of ∼ 125,000 latently infected primary CD4+ T cells after reactivation using three different latency reversing agents. Differentially expressed genes and differentially accessible motifs were used to examine transcriptional pathways and transcription factor (TF) activities across the cell population. We identified cellular transcripts and TFs whose expression/activity was correlated with viral reactivation and demonstrated that a machine learning model trained on these data was 75%–79% accurate at predicting viral reactivation. Finally, we validated the role of two candidate HIV-regulating factors, FOXP1 and GATA3, in viral transcription. These data demonstrate the power of integrated multimodal single-cell analysis to uncover novel relationships between host cell factors and HIV latency.
View publication
Comparison of “framework Shuffling” and “CDR Grafting” in humanization of a PD-1 murine antibody Frontiers in Immunology 2024 Jul

Abstract

IntroductionHumanization is typically adopted to reduce the immunogenicity of murine antibodies generated by hybridoma technology when used in humans.MethodsTwo different strategies of antibody humanization are popularly employed, including “complementarity determining region (CDR) grafting” and “framework (FR) shuffling” to humanize a murine antibody against human programmed death-1 (PD-1), XM PD1. In CDR-grafting humanization, the CDRs of XM PD-1, were grafted into the human FR regions with high homology to the murine FR counterparts, and back mutations of key residues were performed to retain the antigen-binding affinities. While in FR-shuffling humanization, a combinatorial library of the six murine CDRs in-frame of XM PD-1 was constructed to a pool of human germline FRs for high-throughput screening for the most favorable variants. We evaluated many aspects which were important during antibody development of the molecules obtained by the two methods, including antibody purity, thermal stability, binding efficacy, predicted humanness, and immunogenicity, along with T cell epitope prediction for the humanized antibodies.ResultsWhile the ideal molecule was not achieved through CDR grafting in this particular instance, FR-shuffling proved successful in identifying a suitable candidate. The study highlights FR-shuffling as an effective complementary approach that potentially increases the success rate of antibody humanization. It is particularly noted for its accessibility to those with a biological rather than a computational background. DiscussionThe insights from this comparison are intended to assist other researchers in selecting appropriate humanization strategies for drug development, contributing to broader application and understanding in the field.
View publication
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更多信息

更多信息
物种 人类
Magnet Compatibility • EasySep™ Magnet (Catalog #18000) • “The Big Easy” EasySep™ Magnet (Catalog #18001) • Easy 50 EasySep™ Magnet (Catalog #18002) • EasyPlate™ EasySep™ Magnet (Catalog 18102) • EasyEights™ EasySep™ Magnet (Catalog #18103) • RoboSep™-S (Catalog #21000) • Eas
样本来源 PBMC, 白细胞单采术样本
Selection Method Negative
标记抗体
  1. 抗人CD4抗体，clone OKT4
    抗人CD4抗体，clone OKT4

    小鼠Monoclonal IgG2b抗体，抗人、恒河猴、食蟹猴CD4

  2. 抗人CD3抗体，clone UCHT1
    抗人CD3抗体，clone UCHT1

    小鼠（BALB/c）Monoclonal IgG1抗体，抗人、黑猩猩CD3

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