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EasySep™人CD8正选试剂盒 II

人CD8+ T细胞的免疫磁珠正选
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产品号 #（选择产品）

产品号 #17853_C

人CD8+ T细胞的免疫磁珠正选

产品优势

  • 快捷、操作简单
  • 纯度高达99%
  • 无需分离柱     

产品组分包括

  • EasySep™人CD8 正选试剂盒 II（产品号 #17853）
    • EasySep™人CD8正选抗体混合物 II，1 mL
    • EasySep™ Dextran RapidSpheres™ 50100 磁珠，1 mL
    • EasySep™人CD8     正选试剂盒 II（产品号 #100-0699）
    • EasySep™人CD8正选抗体混合物 II，1 x 10 mL
    • EasySep™ Dextran RapidSpheres™ 50103 磁珠，1 x 1 mL
  • RoboSep™人CD8 正选试剂盒 II（产品号 #17853RF）
    • EasySep™人CD8正选抗体混合物 II，1 mL
    • EasySep™ Dextran RapidSpheres™ 50100 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™ 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

总览

使用EasySep™人CD8正选试剂盒 II，可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通过免疫磁珠正选分离出高纯度的人CD8+ T细胞。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™正选过程中，目的细胞经识别CD8的抗体四聚体复合物及磁珠结合标记。抗体复合物中还含有人Fc受体的结合抗体，可最大程度减少非特异性结合。使用EasySep™磁极分离目的细胞，只需简单倾倒或吸出非目的 细胞即可，目的细胞则保留在管中。仅需15分钟磁珠分选后，目的CD8+ T细胞即可用于流式细胞术、细胞培养或DNA/RNA提取等下游应用。CD8抗原表达于  细胞毒性T细胞中，弱表达于NK细胞亚群中     。

该产品可替代EasySep™人CD8细胞正选试剂盒 (产品号 #18053) 以进行更快的细胞分选。

如需从白细胞单采术样本中大规模分选人CD8+细胞，请选用大规格（1 x 10^10细胞）试剂盒（目录号 #100-0699）。

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CD8正选试剂盒 II分离的、符合伦理规范的即用型冻存人原代外周血CD8+ 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™正选过程中，目的细胞经识别CD8的抗体四聚体复合物及磁珠结合标记。抗体复合物中还含有人Fc受体的结合抗体，可最大程度减少非特异性结合。使用EasySep™磁极分离目的细胞，只需简单倾倒或吸出非目的 细胞即可，目的细胞则保留在管中。仅需15分钟磁珠分选后，目的CD8+ T细胞即可用于流式细胞术、细胞培养或DNA/RNA提取等下游应用。CD8抗原表达于  细胞毒性T细胞中，弱表达于NK细胞亚群中     。

该产品可替代EasySep™人CD8细胞正选试剂盒 (产品号 #18053) 以进行更快的细胞分选。

如需从白细胞单采术样本中大规模分选人CD8+细胞，请选用大规格（1 x 10^10细胞）试剂盒（目录号 #100-0699）。

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CD8正选试剂盒 II分离的、符合伦理规范的即用型冻存人原代外周血CD8+ 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™正选过程中，目的细胞经识别CD8的抗体四聚体复合物及磁珠结合标记。抗体复合物中还含有人Fc受体的结合抗体，可最大程度减少非特异性结合。使用EasySep™磁极分离目的细胞，只需简单倾倒或吸出非目的 细胞即可，目的细胞则保留在管中。仅需15分钟磁珠分选后，目的CD8+ T细胞即可用于流式细胞术、细胞培养或DNA/RNA提取等下游应用。CD8抗原表达于  细胞毒性T细胞中，弱表达于NK细胞亚群中     。

该产品可替代EasySep™人CD8细胞正选试剂盒 (产品号 #18053) 以进行更快的细胞分选。

如需从白细胞单采术样本中大规模分选人CD8+细胞，请选用大规格（1 x 10^10细胞）试剂盒（目录号 #100-0699）。

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CD8正选试剂盒 II分离的、符合伦理规范的即用型冻存人原代外周血CD8+ 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™正选过程中，目的细胞经识别CD8的抗体四聚体复合物及磁珠结合标记。抗体复合物中还含有人Fc受体的结合抗体，可最大程度减少非特异性结合。使用EasySep™磁极分离目的细胞，只需简单倾倒或吸出非目的 细胞即可，目的细胞则保留在管中。仅需15分钟磁珠分选后，目的CD8+ T细胞即可用于流式细胞术、细胞培养或DNA/RNA提取等下游应用。CD8抗原表达于  细胞毒性T细胞中，弱表达于NK细胞亚群中     。

该产品可替代EasySep™人CD8细胞正选试剂盒 (产品号 #18053) 以进行更快的细胞分选。

如需从白细胞单采术样本中大规模分选人CD8+细胞，请选用大规格（1 x 10^10细胞）试剂盒（目录号 #100-0699）。

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CD8正选试剂盒 II分离的、符合伦理规范的即用型冻存人原代外周血CD8+ 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) • EasyEights™ EasySep™ Magnet (Catalog #18103) • Easy 50 EasySep™ Magnet (Catalog #18002) • 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，CD8+
 
种属
人
 
样本来源
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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实验数据

Typical EasySep™ Human CD8 Positive Selection Profile

Figure 1. Typical EasySep™ Human CD8 Positive Selection Profile

Starting with a single cell suspension of human PBMCs, the CD8+ cell content of the isolated fraction is typically 96.5 ± 2.4% (mean ± SD) using "The Big Easy" EasySep™ Magnet.

FACS Data for Anti-Human CD8a Antibody, Clone RPA-T8, PerCP-Cy55-Conjugated

Figure 2. FACS Data for Anti-Human CD8a Antibody, Clone RPA-T8, PerCP-Cy55-Conjugated

(A) Flow cytometry analysis of human peripheral blood mononuclear cells (PBMCs) labeled with Anti-Human CD8a Antibody, Clone RPA-T8, PerCP-Cy5.5 (Catalog #60022PS; filled histogram) or a mouse IgG1, kappa isotype control antibody, PerCP-Cy5.5 (solid line histogram).

(B) Flow cytometry analysis of human PBMCs processed with the EasySep™ Human CD8 Positive Selection Kit (Catalog #17853) and labeled with Anti-Human CD8a Antibody, Clone RPA-T8, PerCP-Cy5.5. Histograms show labeling of PBMCs (Start) and isolated cells (Isolated). Labeling of start cells with a mouse IgG1, kappa PerCP-Cy5.5 isotype control antibody is shown (solid line histogram).

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 - Positive Selection
Product Name
RoboSep™ Human CD8 Positive Selection Kit II
Catalog #
17853RF
Lot #
All
Language
Multi
Document Type
Product Information Sheet - Positive Selection
Product Name
RoboSep™ Human CD8 Positive Selection Kit II
Catalog #
17853RF
Lot #
All
Language
English
Document Type
Product Information Sheet - with Easy 250 EasySep™ Magnet
Product Name
EasySep™ Human CD8 Positive Selection Kit II
Catalog #
100-0699
Lot #
All
Language
English
Document Type
Product Information Sheet - Depletion
Product Name
EasySep™ Human CD8 Positive Selection Kit II
Catalog #
17853
Lot #
All
Language
English
Document Type
Product Information Sheet - Positive Selection
Product Name
EasySep™ Human CD8 Positive Selection Kit II
Catalog #
17853
Lot #
All
Language
English
Document Type
Product Information Sheet - Positive Selection
Product Name
EasySep™ Human CD8 Positive Selection Kit II
Catalog #
17853
Lot #
All
Language
Multi
Document Type
Safety Data Sheet 1
Product Name
RoboSep™ Human CD8 Positive Selection Kit II
Catalog #
17853RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Product Name
RoboSep™ Human CD8 Positive Selection Kit II
Catalog #
17853RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 3
Product Name
RoboSep™ Human CD8 Positive Selection Kit II
Catalog #
17853RF
Lot #
All
Language
English
Document Type
Safety Data Sheet
Product Name
EasySep™ Human CD8 Positive Selection Kit II
Catalog #
100-0699
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Product Name
EasySep™ Human CD8 Positive Selection Kit II
Catalog #
17853
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Product Name
EasySep™ Human CD8 Positive Selection Kit II
Catalog #
17853
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 (9)

Isolate Human Immune Cells
Brochure
Isolate Human Immune Cells
EasySep™ Cell Separation Technology
Brochure
EasySep™ Cell Separation Technology
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
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...

常见问题

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 (11)

Fc$\gamma$ receptor-mediated cross-linking codefines the immunostimulatory activity of anti-human CD96 antibodies. A. Rogel et al. JCI insight 2022 oct

Abstract

New strategies that augment T cell responses are required to broaden the therapeutic arsenal against cancer. CD96, TIGIT, and CD226 are receptors that bind to a communal ligand, CD155, and transduce either inhibitory or activating signals. The function of TIGIT and CD226 is established, whereas the role of CD96 remains ambiguous. Using a panel of engineered antibodies, we discovered that the T cell stimulatory activity of anti-CD96 antibodies requires antibody cross-linking and is potentiated by Fc$\gamma$ receptors. Thus, soluble Fc silent" anti-CD96 antibodies failed to stimulate human T cells whereas the same antibodies were stimulatory after coating onto plastic surfaces. Remarkably the activity of soluble anti-CD96 antibodies was reinstated by engineering the Fc domain to a human IgG1 isotype and it was dependent on antibody trans-cross-linking by Fc$\gamma$RI. In contrast neither human IgG2 nor variants with increased Fc$\gamma$ receptor IIB binding possessed stimulatory activity. Anti-CD96 antibodies acted directly on T cells and augmented gene expression networks associated with T cell activation leading to proliferation cytokine secretion and resistance to Treg suppression. Furthermore CD96 expression correlated with survival in HPV+ head and neck squamous cell carcinoma and its cross-linking activated tumor-infiltrating T cells thus highlighting the potential of anti-CD96 antibodies in cancer immunotherapy."
View publication
A genome-scale screen for synthetic drivers of T cell proliferation. M. Legut et al. Nature 2022 mar

Abstract

The engineering of autologous patient T cells for adoptive cell therapies has revolutionized the treatment of several types of cancer1. However, further improvements are needed to increase response and cure rates. CRISPR-based loss-of-function screens have been limited to negative regulators of T cell functions2-4 and raise safety concerns owing to the permanent modification of the genome. Here we identify positive regulators of T cell functions through overexpression of around 12,000 barcoded human open reading frames (ORFs). The top-ranked genes increased the proliferation and activation of primary human CD4+ and CD8+ T cells and their secretion of key cytokines such as interleukin-2 and interferon-$\gamma$. In addition, we developed the single-cell genomics method OverCITE-seq for high-throughput quantification of the transcriptome and surface antigens in ORF-engineered T cells. The top-ranked ORF-lymphotoxin-$\beta$ receptor (LTBR)-is typically expressed in myeloid cells but absent in lymphocytes. When overexpressed in T cells, LTBR induced profound transcriptional and epigenomic remodelling, leading to increased T cell effector functions and resistance to exhaustion in chronic stimulation settings through constitutive activation of the canonical NF-$\kappa$B pathway. LTBR and other highly ranked genes improved the antigen-specific responses of chimeric antigen receptor T cells and ?? T cells, highlighting their potential for future cancer-agnostic therapies5. Our results provide several strategies for improving next-generation T cell therapies by the induction of synthetic cell programmes.
View publication
Chemically modified guide RNAs enhance CRISPR-Cas13 knockdown in human cells. A. M\'endez-Mancilla et al. Cell chemical biology 2022 feb

Abstract

RNA-targeting CRISPR-Cas13 proteins have recently emerged as a powerful platform to modulate gene expression outcomes. However, protein and CRISPR RNA (crRNA) delivery in human cells can be challenging with rapid crRNA degradation yielding transient knockdown. Here we compare several chemical RNA modifications at different positions to identify synthetic crRNAs that improve RNA targeting efficiency and half-life in human cells. We show that co-delivery of modified crRNAs and recombinant Cas13 enzyme in ribonucleoprotein (RNP) complexes can alter gene expression in primary CD4+ and CD8+ T cells. This system represents a robust and efficient method to modulate transcripts without genetic manipulation.
View publication
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更多信息

更多信息
种属 Human
Magnet Compatibility • EasySep™ Magnet (Catalog #18000) • “The Big Easy” EasySep™ Magnet (Catalog #18001) • EasyPlate™ EasySep™ Magnet (Catalog #18102) • EasyEights™ EasySep™ Magnet (Catalog #18103) • Easy 50 EasySep™ Magnet (Catalog #18002) • RoboSep™-S (Catalog #21000) • Ea
样本来源 PBMC
Selection Method Positive
标记抗体
  1. 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

  2. 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

  3. 抗人CD8a抗体，克隆SK1
    抗人CD8a抗体，克隆SK1

    抗人、恒河猴、食蟹猴CD8a的小鼠单克隆IgG1抗体

  4. 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

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