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EasySep™人Pan-DC预富集试剂盒

免疫磁珠负选不带标记的人树突状细胞（包括髓样和浆细胞样树突状细胞）
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产品号 #（选择产品）

产品号 #19251_C

免疫磁珠负选不带标记的人树突状细胞（包括髓样和浆细胞样树突状细胞）

产品优势

  • 操作简单、快捷，且无需分离柱
  • 纯度高达80%
  • 获得不带标记的活细胞

产品组分包括

  • EasySep™人Pan-DC预富集试剂盒（产品号 #19251）
    • EasySep™人Pan-DC预富集抗体混合物组分A，1 mL
    • EasySep™人Pan-DC 预富集抗体混合物组分B，1 mL
    • EasySep™ D Magnetic Particles磁珠，2 x 1 mL
    • 抗人CD32 (Fc gamma RII) 阻断剂，8 mL
  • RoboSep™ 人Pan-DC预富集试剂盒（含过滤吸头）（产品号 #19251RF）
    • EasySep™人Pan-DC预富集抗体混合物组分A，1 mL
    • EasySep™人Pan-DC 预富集抗体混合物组分B，1 mL
    • EasySep™ D Magnetic Particles磁珠，2 x 1 mL
    • 抗人CD32 (Fc gamma RII) 阻断剂，8 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Pan-DC预富集试剂盒，通过免疫磁珠负选，可轻松高效地从新鲜或冻存的人外周血单个核细胞（PBMC）样本中预富集得到高纯度的不带标记的树突状细胞（包括髓样和浆细胞样树突状细胞）。EasySep™技术结合单克隆抗体的特异性和无柱磁分选系统的简便性，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与磁珠标记。以下非目的细胞将被特异性去除：粒细胞、T细胞、B细胞、单核细胞、NK细胞及红系细胞。使用EasySep™磁极吸附后，通过简单地将目的细胞倾倒或吸取至一个新的试管中，即可将被磁珠标记的细胞与不带标记的目的细胞分离开来。磁珠分选后，目的树突细胞可立即用于流式细胞术、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。或者选择用EasySep™人Pan-DC预富集试剂盒分离的即用型、符合伦理来源的原代人外周血冻存树突状细胞。探索更多为您的实验流程优化的产品，包括培养基、补充剂、抗体等。

在该EasySep™负选流程中，非目的细胞会被抗体复合物与磁珠标记。以下非目的细胞将被特异性去除：粒细胞、T细胞、B细胞、单核细胞、NK细胞及红系细胞。使用EasySep™磁极吸附后，通过简单地将目的细胞倾倒或吸取至一个新的试管中，即可将被磁珠标记的细胞与不带标记的目的细胞分离开来。磁珠分选后，目的树突细胞可立即用于流式细胞术、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。或者选择用EasySep™人Pan-DC预富集试剂盒分离的即用型、符合伦理来源的原代人外周血冻存树突状细胞。探索更多为您的实验流程优化的产品，包括培养基、补充剂、抗体等。

在该EasySep™负选流程中，非目的细胞会被抗体复合物与磁珠标记。以下非目的细胞将被特异性去除：粒细胞、T细胞、B细胞、单核细胞、NK细胞及红系细胞。使用EasySep™磁极吸附后，通过简单地将目的细胞倾倒或吸取至一个新的试管中，即可将被磁珠标记的细胞与不带标记的目的细胞分离开来。磁珠分选后，目的树突细胞可立即用于流式细胞术、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。或者选择用EasySep™人Pan-DC预富集试剂盒分离的即用型、符合伦理来源的原代人外周血冻存树突状细胞。探索更多为您的实验流程优化的产品，包括培养基、补充剂、抗体等。

在该EasySep™负选流程中，非目的细胞会被抗体复合物与磁珠标记。以下非目的细胞将被特异性去除：粒细胞、T细胞、B细胞、单核细胞、NK细胞及红系细胞。使用EasySep™磁极吸附后，通过简单地将目的细胞倾倒或吸取至一个新的试管中，即可将被磁珠标记的细胞与不带标记的目的细胞分离开来。磁珠分选后，目的树突细胞可立即用于流式细胞术、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。或者选择用EasySep™人Pan-DC预富集试剂盒分离的即用型、符合伦理来源的原代人外周血冻存树突状细胞。探索更多为您的实验流程优化的产品，包括培养基、补充剂、抗体等。

磁体兼容性
• EasySep™ Magnet (Catalog #18000) • “The Big Easy” EasySep™ Magnet (Catalog #18001) • Easy 50 EasySep™ Magnet (Catalog #18002) • RoboSep™-S (Catalog #21000)
 
亚型
细胞分选试剂盒
 
细胞类型
树突状细胞（DCs）
 
种属
人
 
样本来源
PBMC
 
筛选方法
Negative
 
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 Enrichment Profile For EasySep™ Human Pan-DC Pre-Enrichment Kit

Figure 1. Typical Enrichment Profile For EasySep™ Human Pan-DC Pre-Enrichment Kit

Starting with previously frozen peripheral blood mononuclear cells, the dendritic cell content of the enriched fraction typically ranges from 40% - 80%.

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
Product Name
EasySep™ Human Pan-DC Pre-Enrichment Kit
Catalog #
19251
Lot #
All
Language
English
Document Type
Product Information Sheet
Product Name
RoboSep™ Human Pan-DC Pre-Enrichment Kit with Filter Tips
Catalog #
19251RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Product Name
EasySep™ Human Pan-DC Pre-Enrichment Kit
Catalog #
19251
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Product Name
EasySep™ Human Pan-DC Pre-Enrichment Kit
Catalog #
19251
Lot #
All
Language
English
Document Type
Safety Data Sheet 3
Product Name
EasySep™ Human Pan-DC Pre-Enrichment Kit
Catalog #
19251
Lot #
All
Language
English
Document Type
Safety Data Sheet 4
Product Name
EasySep™ Human Pan-DC Pre-Enrichment Kit
Catalog #
19251
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Product Name
RoboSep™ Human Pan-DC Pre-Enrichment Kit with Filter Tips
Catalog #
19251RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Product Name
RoboSep™ Human Pan-DC Pre-Enrichment Kit with Filter Tips
Catalog #
19251RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 3
Product Name
RoboSep™ Human Pan-DC Pre-Enrichment Kit with Filter Tips
Catalog #
19251RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 4
Product Name
RoboSep™ Human Pan-DC Pre-Enrichment Kit with Filter Tips
Catalog #
19251RF
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)

Cell Enrichment Prior to Cell Sorting
Brochure
Cell Enrichment Prior to Cell Sorting
EasySep™ Cell Separation Technology
Brochure
EasySep™ Cell Separation Technology
Antigen Processing and Presentation
Wallchart
Antigen Processing and Presentation
Human Immune Cytokines
Wallchart
Human Immune Cytokines
Frequencies of Human Cell Types in Blood-Related Sources
Wallchart
Frequencies of Human Cell Types in Blood-Related Sources
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
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
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...
Pre-Enrichment of Dendritic Cells from Human Peripheral Blood Samples
Scientific Poster
Pre-Enrichment of Dendritic Cells from Human Peripheral Blood Samples

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 (4)

Signaling Cascade through DC-ASGPR Induces Transcriptionally Active CREB for IL-10 Induction and Immune Regulation. C. Gu et al. Journal of immunology (Baltimore, Md. : 1950) 2019 jul

Abstract

The types and magnitude of Ag-specific immune responses can be determined by the functional plasticity of dendritic cells (DCs). However, how DCs display functional plasticity and control host immune responses have not been fully understood. In this study, we report that ligation of DC-asialoglycoprotein receptor (DC-ASGPR), a C-type lectin receptor (CLR) expressed on human DCs, resulted in rapid activation of Syk, followed by PLCgamma2 and PKCdelta engagements. However, different from other Syk-coupled CLRs, including Dectin-1, signaling cascade through DC-ASGPR did not trigger NF-kappaB activation. Instead, it selectively activated MAPK ERK1/2 and JNK. Rapid and prolonged phosphorylation of ERK1/2 led to sequential activation of p90RSK and CREB, which consequently bound to IL10 promoter and initiated cytokine expression. In addition, DC-ASGPR ligation activated Akt, which differentially regulated the activities of GSK-3alpha/beta and beta-catenin and further contributed to IL-10 expression. Our observations demonstrate that DC-ASGPR induces IL-10 expression via an intrinsic signaling pathway, which provides a molecular explanation for DC-ASGPR-mediated programing of DCs to control host immune responses.
View publication
Diversification of human plasmacytoid predendritic cells in response to a single stimulus. Alculumbre SG et al. Nature immunology 2018 JAN

Abstract

Innate immune cells adjust to microbial and inflammatory stimuli through a process termed environmental plasticity, which links a given individual stimulus to a unique activated state. Here, we report that activation of human plasmacytoid predendritic cells (pDCs) with a single microbial or cytokine stimulus triggers cell diversification into three stable subpopulations (P1-P3). P1-pDCs (PD-L1+CD80-) displayed a plasmacytoid morphology and specialization for type I interferon production. P3-pDCs (PD-L1-CD80+) adopted a dendritic morphology and adaptive immune functions. P2-pDCs (PD-L1+CD80+) displayed both innate and adaptive functions. Each subpopulation expressed a specific coding- and long-noncoding-RNA signature and was stable after secondary stimulation. P1-pDCs were detected in samples from patients with lupus or psoriasis. pDC diversification was independent of cell divisions or preexisting heterogeneity within steady-state pDCs but was controlled by a TNF autocrine and/or paracrine communication loop. Our findings reveal a novel mechanism for diversity and division of labor in innate immune cells.
View publication
Criteria for Dendritic Cell Receptor Selection for Efficient Antibody-Targeted Vaccination A. Reuter et al. The Journal of Immunology 2015

Abstract

Ab-targeted vaccination involves targeting a receptor of choice expressed by dendritic cells (DCs) with Ag-coupled Abs. Currently, there is little consensus as to which criteria determine receptor selection to ensure superior Ag presentation and immunity. In this study, we investigated parameters of DC receptor internalization and determined how they impact Ag presentation outcomes. First, using mixed bone marrow chimeras, we established that Ag-targeted, but not nontargeted, DCs are responsible for Ag presentation in settings of Ab-targeted vaccination in vivo. Next, we analyzed parameters of DEC205 (CD205), Clec9A, CD11c, CD11b, and CD40 endocytosis and obtained quantitative measurements of internalization speed, surface turnover, and delivered Ag load. Exploiting these parameters in MHC class I (MHC I) and MHC class II (MHC II) Ag presentation assays, we showed that receptor expression level, proportion of surface turnover, or speed of receptor internalization did not impact MHC I or MHC II Ag presentation efficiency. Furthermore, the Ag load delivered to DCs did not correlate with the efficiency of MHC I or MHC II Ag presentation. In contrast, targeting Ag to CD8(+) or CD8(-) DCs enhanced MHC I or MHC II Ag presentation, respectively. Therefore, receptor expression levels, speed of internalization, and/or the amount of Ag delivered can be excluded as major determinants that dictate Ag presentation efficiency in setting of Ab-targeted vaccination.
View publication
View All Publications

更多信息

更多信息
种属 Human
Magnet Compatibility • EasySep™ Magnet (Catalog #18000) • “The Big Easy” EasySep™ Magnet (Catalog #18001) • Easy 50 EasySep™ Magnet (Catalog #18002) • RoboSep™-S (Catalog #21000)
样本来源 PBMC
Selection Method Negative
标记抗体
  1. 抗人CD14抗体，克隆M5E2
    抗人CD14抗体，克隆M5E2

    抗人、恒河猴、食蟹猴CD14的小鼠单克隆IgG2a抗体

  2. 抗人CD16抗体，克隆号3G8
    抗人CD16抗体，克隆号3G8

    小鼠抗人、恒河猴、食蟹猴CD16单克隆IgG1抗体

  3. 抗人CD20抗体，克隆2H7
    抗人CD20抗体，克隆2H7

    小鼠单克隆IgG2b抗体，抗人、恒河猴、食蟹猴CD20

  4. 抗人CD34抗体，克隆581
    抗人CD34抗体，克隆581

    小鼠单克隆IgG1抗体，抗人CD34

  5. 抗人CD56抗体，克隆HCD56
    抗人CD56抗体，克隆HCD56

    小鼠单克隆IgG1抗体，抗人CD56 （NCAM）

  6. 抗人CD123（IL-3Rα）抗体，克隆 6H6
    抗人CD123（IL-3Rα）抗体，克隆 6H6

    抗人、恒河猴、白眉猴CD123 （IL-3Rα）的小鼠单克隆IgG1抗体

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