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）

立即询价

总览

使用EasySep™人CD8正选试剂盒II，通过免疫磁性正选法可从新鲜、冻存的人外周血单个核细胞（PBMCs）或白细胞单采样品中分离高纯度CD8+ T细胞。

该步骤中，CD8+细胞通过识别CD8的抗体复合物和磁珠标记，并使用抗人Fc受体抗体防止非特异结合。标记细胞经EasySep™磁体分离后，未标记细胞被倒出，CD8+细胞保留在管中。整个过程最短仅需15分钟。CD8抗原强表达于细胞毒性T细胞，弱表达于部分NK细胞。

本产品替代旧版EasySep™ CD8试剂盒（产品号#18053），并可扩展至大规模分离（产品号#100-0699）。

实验数据

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.

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。

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
物种	人类
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

EasySep™人CD8正选试剂盒 II

产品号 #100-0699

产品号 #17853

产品号 #17853RF

产品号 #（选择产品）

产品号 #17853_C

产品优势

产品组分包括

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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Scientists Helping Scientists™

EasySep™人CD8正选试剂盒 II

产品号 #100-0699

产品号 #17853

产品号 #17853RF

产品号 #（选择产品）

产品号 #17853_C

产品优势

产品组分包括

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

技术资料 (9)

常见问题

Can EasySep™ be used for either positive or negative selection?

How does the separation work?

Which columns do I use?

How can I analyze the purity of my enriched sample?

Can EasySep™ separations be automated?

Can EasySep™ be used to isolate rare cells?

Are the EasySep™ magnetic particles FACS-compatible?

Can the EasySep™ magnetic particles be removed after enrichment?

Can I alter the separation time in the magnet?

For positive selection, can I perform more than 3 separations to increase purity?

How does the binding of the EasySep™ magnetic particle affect the cells? is the function of positively selected cells altered by the bound particles?

文献 (11)

Abstract

Abstract

Abstract

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更多信息

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