EasySep™ Direct人CD4+ T细胞分选试剂盒

直接从全血中免疫磁珠负选人CD4+ T细胞

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产品号 #（选择产品）

产品号 #19662_C

直接从全血中免疫磁珠负选人CD4+ T细胞

产品优势

> 99.9%的红细胞去除率，无需密度梯度离心、沉降或裂解
分选获得的细胞纯度高达96%
操作简单、快捷，且无需分离柱
分选得到的细胞不带标记

产品组分包括

EasySep™ Direct 人CD4+ T细胞分选试剂盒（产品号 #19662）

EasySep™ Direct人CD4+ T细胞分选抗体混合物, 2 x 2.5 mL
EasySep™ Direct RapidSpheres™ 磁珠, 4 x 2.5 mL

RoboSep™ 人 CD4+ T细胞分选试剂盒（带过滤吸头）（产品号 #19662RF）

EasySep™ Direct人CD4+ T细胞分选抗体混合物, 2 x 2.5 mL
EasySep™ Direct RapidSpheres™ 磁珠, 4 x 2.5 mL
RoboSep™ 缓冲液（产品号 #20104）
RoboSep™过滤吸头（产品号 #20125）x 2

立即询价

总览

使用EasySep™ Direct人 CD4+ T细胞分选试剂盒，可简便高效地从人全血样本中通过免疫磁珠负选获得高纯度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和 EasySep™ Direct RapidSpheres™ 磁珠标记。以下非目的细胞会被特异性去除：粒细胞、CD8+ T细胞和其他非CD4+ T细胞亚群、B细胞、单核细胞、NK细胞和红系细胞。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，目的细胞可通过倾倒或移液吸取轻松收集到新试管中，分选后的细胞可流式细胞术、培养或DNA/RNA提取等下游应用。

了解更多EasySep™免疫磁珠技术的工作原理，或者如何通过RoboSep™实现全自动化免疫磁珠细胞分选。探索更多为您的实验流程优化的产品，包括细胞鉴定、冷冻保存等相关试剂。

实验数据

$Starting with human whole blood from normal healthy donors, the typical CD4+ T cell (CD3+CD4+) content of the non-lysed final isolated fraction is 93.6 ± 2.5% (gated on CD45) or 93.1 ± 2.5% (not gated on CD45).$

Figure 1. Typical EasySep™ Direct Human CD4+ T Cell Isolation Profile

Starting with human whole blood from normal healthy donors, the typical CD4+ T cell (CD3+CD4+) content of the non-lysed final isolated fraction is 93.6 ± 2.5% (gated on CD45) or 93.1 ± 2.5% (not gated on CD45). In the example above, the CD4+ T cell (CD3+CD4+) content of the lysed whole blood start sample and non-lysed final isolated fraction is 16.5% and 95.8% (gated on CD45), respectively, or 16.3% and 95.1% (not gated on CD45), respectively. The starting frequency of CD4+ T cells in the non-lysed whole blood start sample is 0.016% (data not shown).

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 (6)

A genome-wide CRISPR screen identifies a restricted set of HIV host dependency factors. Park RJ et al. Nature genetics 2016 DEC

Abstract

Host proteins are essential for HIV entry and replication and can be important nonviral therapeutic targets. Large-scale RNA interference (RNAi)-based screens have identified nearly a thousand candidate host factors,but there is little agreement among studies and few factors have been validated. Here we demonstrate that a genome-wide CRISPR-based screen identifies host factors in a physiologically relevant cell system. We identify five factors,including the HIV co-receptors CD4 and CCR5,that are required for HIV infection yet are dispensable for cellular proliferation and viability. Tyrosylprotein sulfotransferase 2 (TPST2) and solute carrier family 35 member B2 (SLC35B2) function in a common pathway to sulfate CCR5 on extracellular tyrosine residues,facilitating CCR5 recognition by the HIV envelope. Activated leukocyte cell adhesion molecule (ALCAM) mediates cell aggregation,which is required for cell-to-cell HIV transmission. We validated these pathways in primary human CD4(+) T cells through Cas9-mediated knockout and antibody blockade. Our findings indicate that HIV infection and replication rely on a limited set of host-dispensable genes and suggest that these pathways can be studied for therapeutic intervention.

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Tuning ITAM multiplicity on T cell receptors can control potency and selectivity to ligand density. J. R. James Science signaling 2018 MAY

Abstract

The T cell antigen receptor (TCR) recognizes peptides from pathogenic proteins bound in the major histocompatibility complex (MHC). To convert this binding event into downstream signaling,the TCR complex contains immunoreceptor tyrosine-based activation motifs (ITAMs) that act as docking sites for the cytoplasmic tyrosine kinase ZAP-70. Unique among antigen receptors,the TCR complex uses 10 ITAMs to transduce peptide-MHC binding to the cell interior. Using synthetic,drug-inducible receptor-ligand pairs,it was found that greater ITAM multiplicity primarily enhanced the efficiency with which ligand binding was converted into an intracellular signal. This manifested as an increase in the fraction of cells that became activated in response to antigen,and a more synchronous initiation of TCR-proximal signaling,rather than direct amplification of the intracellular signals. Exploiting these findings,the potency and selectivity of chimeric antigen receptors targeted against cancer were substantially enhanced by modulating the number of encoded ITAMs.

View publication

Multispecific Targeting with Synthetic Ankyrin Repeat Motif Chimeric Antigen Receptors. A. Balakrishnan et al. Clinical cancer research : an official journal of the American Association for Cancer Research 2019 sep

Abstract

PURPOSE The outgrowth of antigen-negative variants is a significant challenge for adoptive therapy with T cells that target a single specificity. Chimeric antigen receptors (CAR) are typically designed with one or two scFvs that impart antigen specificity fused to activation and costimulation domains of T-cell signaling molecules. We designed and evaluated the function of CARs with up to three specificities for overcoming tumor escape using Designed Ankyrin Repeat Proteins (DARPins) rather than scFvs for tumor recognition. EXPERIMENTAL DESIGN A monospecific CAR was designed with a DARPin binder (E01) specific for EGFR and compared with a CAR designed using an anti-EGFR scFv. CAR constructs in which DARPins specific for EGFR,EpCAM,and HER2 were linked together in a single CAR were then designed and optimized to achieve multispecific tumor recognition. The efficacy of CAR-T cells bearing a multispecific DARPin CAR for treating tumors with heterogeneous antigen expression was evaluated in vivo. RESULTS The monospecific anti-EGFR E01 DARPin conferred potent tumor regression against EGFR+ targets that was comparable with an anti-EGFR scFv CAR. Linking three separate DARPins in tandem was feasible and in an optimized format generated a single tumor recognition domain that targeted a mixture of heterogeneous tumor cells,each expressing a single antigen,and displayed synergistic activity when tumor cells expressed more than one target antigen. CONCLUSIONS DARPins can serve as high-affinity recognition motifs for CAR design,and their robust architecture enables linking of multiple binders against different antigens to achieve functional synergy and reduce antigen escape.

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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) • EasyEights™ EasySep™ Magnet (Catalog #18103) • RoboSep™-S (Catalog #21000)
样本来源	全血
Selection Method	Negative

EasySep™ Direct人CD4+ T细胞分选试剂盒

产品号 #19662

产品号 #19662RF

产品号 #（选择产品）

产品号 #19662_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™ Direct人CD4+ T细胞分选试剂盒

产品号 #19662

产品号 #19662RF

产品号 #（选择产品）

产品号 #19662_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 (13)

常见问题 (11)

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 (6)

Abstract

Abstract

Abstract

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