EasySep™人CD14正选试剂盒II

人CD14+细胞的免疫磁珠正选

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产品号 #（选择产品）

产品号 #17858_C

人CD14+细胞的免疫磁珠正选

产品优势

快捷、操作简单
纯度高达97%
无需分离柱

产品组分包括

EasySep™人CD1 4正选试剂盒II（产品号＃17858）

EasySep™人CD14正选抗体混合物II，1mL
EasySep™ Dextran RapidSpheres™ 50100 磁珠,1mL

EasySep™人CD1 4正选试剂盒II（产品号＃100-0694）

EasySep™人CD14正选抗体混合物II,1x10mL
EasySep™ Dextran RapidSpheres™ 50100 磁珠,2x1mL

RoboSep™人CD1 4正选试剂盒II（产品号＃17858RF）

EasySep™人CD14正选抗体混合物II，1mL
EasySep™ Dextran RapidSpheres™ 50100 磁珠,1mL
RoboSep™ 缓冲液（产品号 #20104）
RoboSep™过滤吸头（产品号 #20125）

立即询价

您也可以直接选用即用型、符合伦理来源的总览

使用EasySep™人CD14正选试剂盒II，通过免疫磁正选从新鲜或先冻存的人外周血单个核细胞（PBMCs）或洗净的白细胞样品中分离高纯度的人CD14+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。

在这个EasySep™正选过程中，目的细胞被标记为识别CD14和磁性颗粒的抗体复合物。使用EasySep™磁极和简单地倾倒或移液除去不需要的细胞来分离标记的细胞。感兴趣的细胞留在试管中。在短至22分钟的磁珠分选后，目的CD14+细胞就可以用于流式细胞术、培养或DNA/RNA提取等下游应用。CD14抗原在单核细胞和巨噬细胞上强表达，在粒细胞上弱表达。它也在大多数组织巨噬细胞上表达。

该产品可替代EasySep™人CD14正选试剂盒（产品号#18058）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CD14+细胞，请参见大包装（1x10^10细胞）试剂盒（产品号#100-0694）。

了解更多关于免疫磁珠EasySep™技术工作原理，或如何通过RoboSep™实现全自动化免疫磁珠细胞分选。您也可以直接选用即用型、符合伦理来源的人外周血CD14+单核细胞，冷冻使用EasySep™人CD14正选试剂盒II进行分选。探索更多为您实验流程优化的产品，包括培养基、添加剂、抗体等。

实验数据

Figure 1. Typical EasySep™ Human CD14 Positive Selection II Isolation Profile

Starting with a single cell suspension of human PBMCs, the CD14+ cell content of the isolated fraction is typically 95.3 ± 4.5% (mean ± SD using the purple EasySep™ Magnet).

Figure 2. FACS Data for Anti-Human CD14 Antibody, Clone M5E2, Alexa Fluor® 488-Conjugated

(A) Flow cytometry analysis of human peripheral blood mononuclear cells (PBMCs) labeled with Anti-Human CD14 Antibody, Clone M5E2, Alexa Fluor® 488 (Catalog #60004AD) and Anti-Human CD45 Antibody, Clone HI30, APC (Catalog #60018AD). (B) Flow cytometry analysis of human PBMCs processed with the EasySep™ Human CD14 Positive Selection Kit (Catalog #17858) and labeled with Anti-Human CD14 Antibody, Clone M5E2, Alexa Fluor® 488. Histograms show labeling of PBMCs (Start) and isolated cells (Isolated). Labeling of start cells with Mouse IgG2a, kappa Isotype Control Antibody, Clone MOPC-173, Alexa Fluor® 488 (Catalog #60071AD) is shown (solid line histogram). (C) Flow cytometry analysis of human whole blood nucleated cells processed with the EasySep™ HLA Whole Blood CD33 Positive Selection Kit (Catalog #17885) and labeled with Anti-Human CD14 Antibody, Clone M5E2, Alexa Fluor® 488. Histograms show labeling of whole blood nucleated cells (Start) and isolated cells (Isolated). Labeling of start cells with Mouse IgG2a, kappa Isotype Control Antibody, Clone MOPC-173, Alexa Fluor® 488 is shown (solid line histogram).

Figure 3. FACS Data for Anti-Human CD14 Antibody, Clone M5E2, PE-Conjugated

(A) Flow cytometry analysis of human peripheral blood mononuclear cells (PBMCs) labeled with Anti-Human CD14 Antibody, Clone M5E2, PE (filled histogram; Catalog #60004PE), or Mouse IgG2a, kappa Isotype Control Antibody, Clone HI30, APC (Catalog #60018AZ). (B) Flow cytometry analysis of human PBMCs processed with the EasySep™ Human CD14 Positive Selection Kit (Catalog #17858) and labeled with Anti-Human CD14 Antibody, Clone M5E2, PE. Histograms show labeling of PBMCs (Start) and isolated cells (Isolated). Labeling of start cells with Mouse IgG2a, kappa Isotype Control Antibody, Clone MOPC-173, PE (Catalog #60071PE) is shown (solid line historgram). (C) Flow cytometry analysis of human whole blood nucleated cells processed with the EasySep™ HLA Whole Blood CD33 Positive Selection Kit (Catalog #17885) and labeled with Anti-Human CD14 Antibody, Clone M5E2, FITC. Histograms show labeling of whole blood nucleated cells (Start) and isolated cells (Isolated). Labeling of start cells with Mouse IgG2a, kappa Isotype Control Antibody, Clone MOPC-173, PE 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。

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。

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 (43)

IFN-Stimulated Gene LY6E in Monocytes Regulates the CD14/TLR4 Pathway but Inadequately Restrains the Hyperactivation of Monocytes during Chronic HIV-1 Infection Xu X et al. The Journal of Immunology 2014

Abstract

Owing to ongoing recognition of pathogen-associated molecular patterns,immune activation and upregulation of IFN-stimulated genes (ISGs) are sustained in the chronically infected host. Albeit most ISGs are important effectors for containing viral replication,some might exert compensatory immune suppression to limit pathological dysfunctions,although the mechanisms are not fully understood. In this study,we report that the ISG lymphocyte Ag 6 complex,locus E (LY6E) is a negative immune regulator of monocytes. LY6E in monocytes negatively modulated CD14 expression and subsequently dampened the responsiveness to LPS stimulation in vitro. In the setting of chronic HIV infection,the upregulation of LY6E was correlated with reduced CD14 level on monocytes; however,the immunosuppressive effect of LY6E was not adequate to remedy the hyperresponsiveness of activated monocytes. Taken together,the regulatory LY6E pathway in monocytes represents one of negative feedback mechanisms that counterbalance monocyte activation,which might be caused by LPS translocation through the compromised gastrointestinal tract during persistent HIV-1 infection and may serve as a potential target for immune intervention.

View publication

Genetic Vaccines To Potentiate the Effective CD103+ Dendritic Cell-Mediated Cross-Priming of Antitumor Immunity Y. Zhang et al. The Journal of Immunology 2015

Abstract

The development of effective cancer vaccines remains an urgent,but as yet unmet,clinical need. This deficiency is in part due to an incomplete understanding of how to best invoke dendritic cells (DC) that are crucial for the induction of tumor-specific CD8(+) T cells capable of mediating durable protective immunity. In this regard,elevated expression of the transcription factor X box-binding protein 1 (XBP1) in DC appears to play a decisive role in promoting the ability of DC to cross-present Ags to CD8(+) T cells in the therapeutic setting. Delivery of DNA vaccines encoding XBP1 and tumor Ag to skin DC resulted in increased IFN-? production by plasmacytoid DC (pDC) from skin/tumor draining lymph nodes and the cross-priming of Ag-specific CD8(+) T cell responses associated with therapeutic benefit. Antitumor protection was dependent on cross-presenting Batf3(+) DC,pDC,and CD8(+) T cells. CD103(+) DC from the skin/tumor draining lymph nodes of the immunized mice appeared responsible for activation of Ag-specific naive CD8(+) T cells,but were dependent on pDC for optimal effectiveness. Similarly,human XBP1 improved the capacity of human blood- and skin-derived DC to activate human T cells. These data support an important intrinsic role for XBP1 in DC for effective cross-priming and orchestration of Batf3(+) DC-pDC interactions,thereby enabling effective vaccine induction of protective antitumor immunity.

View publication

Staphylococcus aureus-derived factors induce IL-10, IFN-γ and IL-17A-expressing FOXP3(+)CD161(+) T-helper cells in a partly monocyte-dependent manner. Bjö et al. Scientific Reports 2016 FEB

Abstract

Staphylococcus aureus (S. aureus) is a human pathogen as well as a frequent colonizer of skin and mucosa. This bacterium potently activates conventional T-cells through superantigens and it is suggested to induce T-cell cytokine-production as well as to promote a regulatory phenotype in T-cells in order to avoid clearance. This study aimed to investigate how S. aureus impacts the production of regulatory and pro-inflammatory cytokines and the expression of CD161 and HELIOS by peripheral CD4(+)FOXP3(+) T-cells. Stimulation of PBMC with S. aureus 161:2-cell free supernatant (CFS) induced expression of IL-10,IFN-γ and IL-17A in FOXP3(+) cells. Further,CD161 and HELIOS separated the FOXP3(+) cells into four distinct populations regarding cytokine-expression. Monocyte-depletion decreased S. aureus 161:2-induced activation of FOXP3(+) cells while pre-stimulation of purified monocytes with S. aureus 161:2-CFS and subsequent co-culture with autologous monocyte-depleted PBMC was sufficient to mediate activation of FOXP3(+) cells. Together,these data show that S. aureus potently induces FOXP3(+) cells and promotes a diverse phenotype with expression of regulatory and pro-inflammatory cytokines connected to increased CD161-expression. This could indicate potent regulation or a contribution of FOXP3(+) cells to inflammation and repression of immune-suppression upon encounter with S. aureus.

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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) • 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CD14正选试剂盒II

产品号 #17858

产品号 #17858RF

产品号 #100-0694

产品号 #（选择产品）

产品号 #17858_C

产品优势

产品组分包括

您也可以直接选用即用型、符合伦理来源的总览

实验数据

产品说明书及文档

应用领域

相关材料与文献

Request Pricing

更多信息

Scientists Helping Scientists™

EasySep™人CD14正选试剂盒II

产品号 #17858

产品号 #17858RF

产品号 #100-0694

产品号 #（选择产品）

产品号 #17858_C

产品优势

产品组分包括

您也可以直接选用即用型、符合伦理来源的总览

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 (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 (43)

Abstract

Abstract

Abstract

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