概述

EasySep™人γ / δ T细胞分离试剂盒使用EasySep™人γ / δ T细胞分离试剂盒，通过免疫磁阴性选择，从新鲜或先前冷冻的人外周血单个核细胞（PBMCs）或裂解的白细胞分离样品中轻松高效地分离高度纯化的人γ / δ T细胞。EasySep™在已发表的研究中广泛使用了20多年，它结合了单克隆抗体的特异性和无柱磁系统的简单性。

在这个EasySep™阴性选择程序中，用抗体复合物和磁性颗粒标记不需要的细胞（包括CD16和CD25细胞）。然后使用EasySep™磁铁将磁性标记的细胞与未接触的所需γ / δ T细胞分离，并将所需细胞倒入或移液到新管中。磁性细胞分离后，所需的γ / δ T细胞可用于流式细胞术、培养或DNA/RNA提取等下游应用。

了解更多关于免疫磁性的知识EasySep™技术工作原理或如何完全自动化免疫磁细胞分离RoboSep™．或者，选择现成的、道德来源的、主要的人外周血γ δ T细胞，冷冻使用EasySep™人γ / δ T细胞分离试剂盒进行分离。探索额外的产品针对您的工作流程进行了优化，包括培养基、补充剂、抗体等。

Magnet Compatibility

• EasySep™ Magnet (Catalog #18000)

• “The Big Easy” EasySep™ Magnet (Catalog #18001)

• Easy 50 EasySep™ Magnet (Catalog #18002)

• RoboSep™-S (Catalog #21000)

Subtype
Cell Isolation Kits

Cell Type
T Cells, T Cells, Other Subsets

Species
Human

Sample Source
Leukapheresis, PBMC

Selection Method
Negative

Application
Cell Isolation

Brand
EasySep, RoboSep

Area of Interest
Immunology

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Data Figures

Figure 1. Typical EasySep™ Human Gamma/Delta T Cell Isolation Profile

Starting with fresh PBMCs, the gamma/delta TCR+CD3+ cell content of the isolated fraction typically ranges from 90 - 97%. In the above example, the purities of the start and final isolated fractions are 2.5% and 92.9%, respectively.

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 Gamma/Delta T Cell Isolation Kit

Catalog #

19255

Lot #

All

Language

English

Document Type

Product Information Sheet

Product Name

RoboSep™ Human Gamma/Delta T Cell Isolation Kit

Catalog #

19255RF

Lot #

All

Language

English

Document Type

Safety Data Sheet 1

Product Name

EasySep™ Human Gamma/Delta T Cell Isolation Kit

Catalog #

19255

Lot #

All

Language

English

Document Type

Safety Data Sheet 2

Product Name

EasySep™ Human Gamma/Delta T Cell Isolation Kit

Catalog #

19255

Lot #

All

Language

English

Document Type

Safety Data Sheet 1

Product Name

RoboSep™ Human Gamma/Delta T Cell Isolation Kit

Catalog #

19255RF

Lot #

All

Language

English

Document Type

Safety Data Sheet 2

Product Name

RoboSep™ Human Gamma/Delta T Cell Isolation Kit

Catalog #

19255RF

Lot #

All

Language

English

Document Type

Safety Data Sheet 3

Product Name

RoboSep™ Human Gamma/Delta T Cell Isolation Kit

Catalog #

19255RF

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

Workflow Stages for T Cell Research

Cell Sourcing

Cell Isolation

Cell Activation, Expansion, and Differentiation

Cell Characterization

Workflow Stages for T Cell Engineering

Cell Isolation

Cell Activation, Expansion, and Differentiation

Cell Characterization

Cell Cryopreservation

Resources and Publications

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.

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.

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1$\alpha$,25(OH)2D3 reverses exhaustion and enhances antitumor immunity of human cytotoxic T cells. P. Li et al. Journal for immunotherapy of cancer 2022 mar

Abstract

BACKGROUND Epidemiological surveys have revealed that low serum vitamin D level was correlated with increased risk of tumors. Dysfunctional T cells in patients with tumor are characterized as exhausted with high levels of immune checkpoint receptors (ICRs). However, whether the reduced level of vitamin D in patients with cancer correlates with cytotoxic T-cell exhaustion is unknown. METHODS Periphery blood samples from 172 patients with non-small cell lung cancer (NSCLC) were prospectively collected. Patients with NSCLC received one course of intravenous docetaxel (75 mg/m2) followed by treatment with or without rocaltrol at a dose of 0.5-2.0 µg/day for total of 3 weeks. We performed phenotypical and functional analysis of T-cell through flow cytometry. Vitamin D receptor (VDR) knockout and overexpression CD8+ and V$\delta$2+ T cells were constructed using Cas9-gRNA targeted and overexpressing approaches to identify 1$\alpha$,25(OH)2D3/VDR-mediated transcription regulation for ICRs or antitumor activity in T cells. RESULTS We show that serum level of vitamin D is negatively correlated with expression of programmed cell death-1 (PD-1), T-cell immunoreceptor with Ig and ITIM domains (TIGIT), and T-cell immunoglobulin and mucin-domain containing-3 (Tim-3), but positively correlated with CD28 expression on CD8+ and V$\gamma$9V$\delta$2+ T cells in patients with NSCLC. 1$\alpha$,25(OH)2D3, the active form of vitamin D, promotes the nuclear translocation of VDR, which binds to the promoter region of Pdcd1, Tim3, and Tigit genes and inhibits their expression. Besides, 1$\alpha$,25(OH)2D3 pretreatment also promotes the methylation of CpG island in the promoter region of the Pdcd1 gene and increases H3K27 acetylation at the promoter region of the Cd28 gene, which leads to surface PD-1 downregulation and CD28 upregulation, respectively. We further reveal that VDR-mediated Ca2+ influx enhanced expression of Th1 cytokines via T-cell receptor activation. Functionally, 1$\alpha$,25(OH)2D3 pretreated CD8+ T cells or V$\gamma$9V$\delta$2+ T cells showed increased Th1 cytokine production and enhanced antitumor immunity. Finally, oral 1$\alpha$,25(OH)2D3 could also decrease expression of PD-1, Tim-3, TIGIT and increase expression of CD28, resulting in cytokine production (associated with antitumor immunity) by cytotoxic T cells of patients with NSCLC. CONCLUSIONS Our findings uncover the pleiotropic effects of 1$\alpha$,25(OH)2D3 in rescuing the exhausted phenotype of human cytotoxic T cells in patients with tumor and in promoting their antitumor immunity. TRIAL REGISTRATION NUMBER ChiCTR2100051135.

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Targeted glycan degradation potentiates the anticancer immune response in vivo. M. A. Gray et al. Nature chemical biology 2020 dec

Abstract

Currently approved immune checkpoint inhibitor therapies targeting the PD-1 and CTLA-4 receptor pathways are powerful treatment options for certain cancers; however, most patients across cancer types still fail to respond. Consequently, there is interest in discovering and blocking alternative pathways that mediate immune suppression. One such mechanism is an upregulation of sialoglycans in malignancy, which has been recently shown to inhibit immune cell activation through multiple mechanisms and therefore represents a targetable glycoimmune checkpoint. Since these glycans are not canonically druggable, we designed an $\alpha$HER2 antibody-sialidase conjugate that potently and selectively strips diverse sialoglycans from breast cancer cells. In syngeneic breast cancer models, desialylation enhanced immune cell infiltration and activation and prolonged the survival of mice, an effect that was dependent on expression of the Siglec-E checkpoint receptor found on tumor-infiltrating myeloid cells. Thus, antibody-sialidase conjugates represent a promising modality for glycoimmune checkpoint therapy.

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EasySep™人γ / δ T细胞分离试剂盒

产品号 #（选择产品）

产品号 #19255_C

产品优势

产品组分包括

概述

Data Figures

Protocols and Documentation

Applications

Resources and Publications

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Scientists Helping Scientists™

Species	Human
Magnet Compatibility	• EasySep™ Magnet (Catalog #18000) • “The Big Easy” EasySep™ Magnet (Catalog #18001) • Easy 50 EasySep™ Magnet (Catalog #18002) • RoboSep™-S (Catalog #21000)
Sample Source	Leukapheresis, PBMC
Selection Method	Negative

EasySep™人γ / δ T细胞分离试剂盒

产品号 #（选择产品）

产品号 #19255_C

产品优势

产品组分包括

概述

Data Figures

Protocols and Documentation

Applications

Resources and Publications

Educational Materials (8)

Frequently Asked Questions

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?

Publications (3)

Abstract

Abstract

Abstract

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