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EasySep™人CD8+ T细胞分选试剂盒

人CD8+ T细胞的免疫磁珠负选
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产品号 #（选择产品）

产品号 #17953_C

人CD8+ T细胞的免疫磁珠负选

产品优势

  • 快捷、操作简单，且无需分离柱
  • 纯度高达91%，回收率高
  • 获得的活细胞无标记

产品组分包括

  • EasySep™人CD8+ T细胞分选试剂盒（产品号 #17953）
    • EasySep™人CD8+ T细胞分选抗体混合物，1 mL
    • EasySep™ Dextran RapidSpheres™磁珠，1 mL
  • EasySep™人CD8+ T细胞分选试剂盒（产品号 #100-0710）
    • EasySep™人CD8+ 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CD8+ T细胞分选试剂盒（产品号 #17953RF）
    • EasySep™人CD8+ 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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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 CD8⁺ T 细胞分选试剂盒，通过免疫磁性负选，可轻松高效地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样本中分离高纯度的人 CD8⁺ T 细胞。EasySep™ 技术结合单克隆抗体的特异性与无柱磁系统的简便性，已在发表的研究中广泛应用超过 20 年。   

在此 EasySep™ 负选步骤中，不需要的细胞通过抗体复合物和磁珠标记。表达以下标记的非目的细胞将被去除：CD4、CD14、CD16、CD19、CD20、CD36、CD56、CD66b、CD123、GlyA 和 TCRγδ。使用 EasySep™ 磁极进行无柱分选，磁珠标记的细胞被去除，只需将未标记的CD8⁺ T 细胞倒入或移取至新的管中即可。仅需 8 分钟即可完成磁珠分选过程，分离后的CD8⁺ T细胞可直接用于下游应用，例如流式细胞术、细胞培或DNA/RNA 提取。

本产品可替代 EasySep™ 人 CD8⁺ T 细胞富集试剂盒（产品号 #19053），实现更快速的细胞分选。

如需从白细胞单采术样本中进行大规模（1×1010 个细胞）的人 CD8⁺ T 细胞分离，请参阅大规格试剂盒（产品号 #100-0710）。

了解更多关于 EasySep™ 免疫磁性技术的工作原理，或了解如何使用 RoboSep™ 实现全自动化的免疫磁珠细胞分选。您也可以选择使用 EasySep™ 人 CD8⁺ T 细胞分选试剂盒制备的即用型、符合伦理的冻存人外周血 CD8⁺ T 细胞。探索更多优化实验流程的产品，包括培养基、补充物、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，CD8+
 
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

EasySep™ Human CD8+ T Cell Isolation Kit

Figure 1. EasySep™ Human CD8+ T Cell Isolation Kit

Starting with human peripheral blood mononuclear cells (PBMCs), the CD8+ T cell content (CD3+CD8+) of the isolated fraction is typically 85.6 ± 4.9% (mean ± SD for the purple EasySep™ Magnet).

Experimental Workflow: DC/T Cell Co-Culture Protocol for the Activation and Expansion of Antigen-Induced CD8+ T Cells

Figure 2. Experimental Workflow: DC/T Cell Co-Culture Protocol for the Activation and Expansion of Antigen-Induced CD8+ T Cells

The EasySep™ Human CD8+ T Cell Isolation Kit (Catalog #17953) can be used as a part of a workflow to assess antigen-specific T cell functionality by co-culturing dendritic cells (DCs) and CD8+ T cells. (1) Isolate monocytes from Human Peripheral Blood Leukopak, Fresh (Catalog #70500) or from Human Peripheral Blood Mononuclear Cells (PBMCs), Fresh or Frozen (Catalog #70025), using EasySep™ Human Monocyte Isolation Kit (Catalog #19359). (2) Culture monocytes to generate monocyte-derived dendritic cells (Mo-DCs) using ImmunoCult™ Dendritic Cell Culture Kit (Catalog #10985) and the peptide(s) of interest. (3) Isolate CD8+ T cells from the same donor’s blood or PBMCs using EasySep™ Human CD8+ T Cell Isolation Kit. (4) Co-culture DCs and CD8+ T cells in ImmunoCult™-XF T Cell Expansion Medium (Catalog #10981). (5) For short-term co-culture, isolate CD8+ T cells using EasySep™ Human CD8+ Cell Isolation Kit and label the isolated CD8+ T cells with a cell proliferation tracking dye. Set up co-culture by seeding peptide-pulsed dendritic cell suspension with the CD8+ T cell suspension at a 1:4 ratio. Harvest the co-cultures and quantify the antigen-specific CD8+ T cells with tetramer staining after 6 days. (6) For long-term co-culture, expand antigen-specific CD8+ T cells with additional supplements. Analyze the phenotype and function of expanded CD8+ T cells by assessing surface markers or cytokine production. Alternatively, enrich antigen-specific CD8+ T cells with EasySep™, rest the cells for 2 days, and then assess killing activity.

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
产品说明书
Product Name
RoboSep™ 人CD8+ T细胞分选试剂盒
Catalog #
17953RF
Lot #
All
Language
中文
Document Type
产品说明书
Product Name
EasySep™人CD8+ T细胞分选试剂盒
Catalog #
17953
Lot #
All
Language
中文
Document Type
Product Information Sheet - with Easy 250 EasySep™ Magnet
Product Name
EasySep™ Human CD8+ T Cell Isolation Kit
Catalog #
100-0710
Lot #
All
Language
English
Document Type
Product Information Sheet
Product Name
EasySep™ Human CD8+ T Cell Isolation Kit
Catalog #
17953
Lot #
All
Language
English
Document Type
Product Information Sheet
Product Name
RoboSep™ Human CD8+ T Cell Isolation Kit
Catalog #
17953RF
Lot #
All
Language
English
Document Type
Safety Data Sheet
Product Name
EasySep™ Human CD8+ T Cell Isolation Kit
Catalog #
100-0710
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Product Name
EasySep™ Human CD8+ T Cell Isolation Kit
Catalog #
17953
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Product Name
EasySep™ Human CD8+ T Cell Isolation Kit
Catalog #
17953
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Product Name
RoboSep™ Human CD8+ T Cell Isolation Kit
Catalog #
17953RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Product Name
RoboSep™ Human CD8+ T Cell Isolation Kit
Catalog #
17953RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 3
Product Name
RoboSep™ Human CD8+ T Cell Isolation Kit
Catalog #
17953RF
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 (17)

CAR-T细胞的制备
挂图
CAR-T细胞的制备
Cell Enrichment Prior to Cell Sorting
产品手册
Cell Enrichment Prior to Cell Sorting
EasySep™细胞分选技术 - 更高效地分离细胞
产品手册
EasySep™细胞分选技术 - 更高效地分离细胞
Isolate Human Immune Cells
产品手册
Isolate Human Immune Cells
细胞分选相关工具
产品手册
细胞分选相关工具
Dendritic Cell/CD8+ T Cell Co-Culture to Assess Antigen-Specific T Cell Functionality
技术公告
Dendritic Cell/CD8+ T Cell Co-Culture to Assess Antigen-Specific T Cell Functionality
Isolate Cells with a Simple Pour-Off: EasySep™ Cell Separation Technology
视频
Isolate Cells with a Simple Pour-Off: EasySep™ Cell Separation Technology
How to Isolate PBMCs from Whole Blood Using Density Gradient Centrifugation (Ficoll™ or Lymphoprep™)
视频
How to Isolate PBMCs from Whole Blood Using Density Gradient Centrifugation (Ficoll™ or Lymphoprep™)
How to Isolate Cells in 96-Well Plates Using the EasyPlate™ Cell Separation Magnet
视频
How to Isolate Cells in 96-Well Plates Using the EasyPlate™ Cell Separation Magnet
Simultaneous Cell Isolation from Multiple Samples Using the EasyEights™ EasySep™ Magnet
视频
Simultaneous Cell Isolation from Multiple Samples Using the EasyEights™ EasySep™ Magnet
How to Isolate Cells with EasySep™ Column-Free Cell Separation Technology
视频
How to Isolate Cells with EasySep™ Column-Free Cell Separation Technology
How EasySep™ Magnetic Cell Separation Technology Works: Fast and Easy Cell Isolation
视频
How EasySep™ Magnetic Cell Separation Technology Works: Fast and Easy Cell Isolation
T Cell Differentiation and Cancer Immunity
视频
T Cell Differentiation and Cancer Immunity
Human Immune Cytokines
挂图
Human Immune Cytokines
Antigen Processing and Presentation
挂图
Antigen Processing and Presentation
Next-Generation Tools to Advance Your Human T Cell Research
线上讲座
Next-Generation Tools to Advance Your Human T Cell Research
Hematopoietic Stem Cell Fitness and Function During Sickle Cell Disease
线上讲座
Hematopoietic Stem Cell Fitness and Function During Sickle Cell Disease

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 (24)

Expression profiling of human immune cell subsets identifies miRNA-mRNA regulatory relationships correlated with cell type specific expression. Allantaz F et al. PloS one 2012

Abstract

Blood consists of different cell populations with distinct functions and correspondingly,distinct gene expression profiles. In this study,global miRNA expression profiling was performed across a panel of nine human immune cell subsets (neutrophils,eosinophils,monocytes,B cells,NK cells,CD4 T cells,CD8 T cells,mDCs and pDCs) to identify cell-type specific miRNAs. mRNA expression profiling was performed on the same samples to determine if miRNAs specific to certain cell types down-regulated expression levels of their target genes. Six cell-type specific miRNAs (miR-143; neutrophil specific,miR-125; T cells and neutrophil specific,miR-500; monocyte and pDC specific,miR-150; lymphoid cell specific,miR-652 and miR-223; both myeloid cell specific) were negatively correlated with expression of their predicted target genes. These results were further validated using an independent cohort where similar immune cell subsets were isolated and profiled for both miRNA and mRNA expression. miRNAs which negatively correlated with target gene expression in both cohorts were identified as candidates for miRNA/mRNA regulatory pairs and were used to construct a cell-type specific regulatory network. miRNA/mRNA pairs formed two distinct clusters in the network corresponding to myeloid (nine miRNAs) and lymphoid lineages (two miRNAs). Several myeloid specific miRNAs targeted common genes including ABL2,EIF4A2,EPC1 and INO80D; these common targets were enriched for genes involved in the regulation of gene expression (ptextless9.0E-7). Those miRNA might therefore have significant further effect on gene expression by repressing the expression of genes involved in transcriptional regulation. The miRNA and mRNA expression profiles reported in this study form a comprehensive transcriptome database of various human blood cells and serve as a valuable resource for elucidating the role of miRNA mediated regulation in the establishment of immune cell identity.
View publication
Self-reactive IgE exacerbates interferon responses associated with autoimmunity. Henault J et al. Nature Immunology 2016 FEB

Abstract

Canonically,immunoglobulin E (IgE) mediates allergic immune responses by triggering mast cells and basophils to release histamine and type 2 helper cytokines. Here we found that in human systemic lupus erythematosus (SLE),IgE antibodies specific for double-stranded DNA (dsDNA) activated plasmacytoid dendritic cells (pDCs),a type of cell of the immune system linked to viral defense,which led to the secretion of substantial amounts of interferon-α (IFN-α). The concentration of dsDNA-specific IgE found in patient serum correlated with disease severity and greatly potentiated pDC function by triggering phagocytosis via the high-affinity FcɛRI receptor for IgE,followed by Toll-like receptor 9 (TLR9)-mediated sensing of DNA in phagosomes. Our findings expand the known pathogenic mechanisms of IgE-mediated inflammation beyond those found in allergy and demonstrate that IgE can trigger interferon responses capable of exacerbating self-destructive autoimmune responses.
View publication
Melanoma cell lysosome secretory burst neutralizes the CTL-mediated cytotoxicity at the lytic synapse. Khazen R et al. Nature Communications 2016 MAR

Abstract

Human melanoma cells express various tumour antigens that are recognized by CD8(+) cytotoxic T lymphocytes (CTLs) and elicit tumour-specific responses in vivo. However,natural and therapeutically enhanced CTL responses in melanoma patients are of limited efficacy. The mechanisms underlying CTL effector phase failure when facing melanomas are still largely elusive. Here we show that,on conjugation with CTL,human melanoma cells undergo an active late endosome/lysosome trafficking,which is intensified at the lytic synapse and is paralleled by cathepsin-mediated perforin degradation and deficient granzyme B penetration. Abortion of SNAP-23-dependent lysosomal trafficking,pH perturbation or impairment of lysosomal proteolytic activity restores susceptibility to CTL attack. Inside the arsenal of melanoma cell strategies to escape immune surveillance,we identify a self-defence mechanism based on exacerbated lysosome secretion and perforin degradation at the lytic synapse. Interfering with this synaptic self-defence mechanism might be useful in potentiating CTL-mediated therapies in melanoma patients.
View publication
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更多信息

更多信息
物种 人
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CD8a抗体，clone RPA-T8
    抗人CD8a抗体，clone RPA-T8

    小鼠Monoclonal IgG1抗体，抗人、恒河猴、食蟹猴CD8a

  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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