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人全血CD34+细胞分选完全试剂盒

免疫磁珠正选
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产品号 #（选择产品）

产品号 #15086_C

免疫磁珠正选

产品优势

  • 易于操作、快速
  • 纯度高达98%
  • 无需分离柱
  • 可与SepMate™联合使用，实现一致且高通量的样本处理

产品组分包括

  • RoboSep™人全血CD34分选完全试剂盒 (产品号15086RF)
    • RosetteSep™人造血祖细胞富集抗体混合物II，3 x 2 mL
    • EasySep™人CD34正选抗体混合物，0.4 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）
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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. Lymphoprep™
    Lymphoprep™

    Density gradient medium for the isolation of mononuclear cells

  2. Dulbecco's Phosphate Buffered Saline with 2% Fetal Bovine Serum
    Dulbecco's Phosphate Buffered Saline with 2% ...

    Cell culture buffer

  3. The Big Easy EasySep™ Magnet
    "The Big Easy" EasySep™ Magnet

    Magnet for column-free immunomagnetic separation

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

人全血CD34+细胞完全试剂盒是一种简单的两步法，用于从全血或白膜层中分离CD34+细胞。

首先，使用RosetteSep™人造血祖细胞富集抗体混合物 (产品号 #15186C)预富集造血祖细胞，包含可识别CD2, CD3, CD14, CD16, CD19, CD24, CD56, CD66b和CD61细胞表面标志物的抗体。然后使用EasySep™人CD34正选抗体混合物 (产品号 #18066C) 来分选CD34+细胞，其中包含识别CD34的抗体。RosetteSep™将非目的细胞与红细胞 (RBCs) 交联，形成免疫玫瑰花环，并在密度梯度离心时沉降。含CD34+细胞的预富集组分可从血浆和密度梯度离心液介质的界面收集。预富集的CD34+细胞随后用抗体和磁珠标记，并通过EasySep™磁极，无需分离柱分选。非目的细胞通过简单倾倒弃去，而目的细胞则保留在管中。分选后的CD34+细胞可立即用于下游应用。

首先，使用RosetteSep™人造血祖细胞富集抗体混合物 (产品号 #15186C)预富集造血祖细胞，包含可识别CD2, CD3, CD14, CD16, CD19, CD24, CD56, CD66b和CD61细胞表面标志物的抗体。然后使用EasySep™人CD34正选抗体混合物 (产品号 #18066C) 来分选CD34+细胞，其中包含识别CD34的抗体。RosetteSep™将非目的细胞与红细胞 (RBCs) 交联，形成免疫玫瑰花环，并在密度梯度离心时沉降。含CD34+细胞的预富集组分可从血浆和密度梯度离心液介质的界面收集。预富集的CD34+细胞随后用抗体和磁珠标记，并通过EasySep™磁极，无需分离柱分选。非目的细胞通过简单倾倒弃去，而目的细胞则保留在管中。分选后的CD34+细胞可立即用于下游应用。

首先，使用RosetteSep™人造血祖细胞富集抗体混合物 (产品号 #15186C)预富集造血祖细胞，包含可识别CD2, CD3, CD14, CD16, CD19, CD24, CD56, CD66b和CD61细胞表面标志物的抗体。然后使用EasySep™人CD34正选抗体混合物 (产品号 #18066C) 来分选CD34+细胞，其中包含识别CD34的抗体。RosetteSep™将非目的细胞与红细胞 (RBCs) 交联，形成免疫玫瑰花环，并在密度梯度离心时沉降。含CD34+细胞的预富集组分可从血浆和密度梯度离心液介质的界面收集。预富集的CD34+细胞随后用抗体和磁珠标记，并通过EasySep™磁极，无需分离柱分选。非目的细胞通过简单倾倒弃去，而目的细胞则保留在管中。分选后的CD34+细胞可立即用于下游应用。

首先，使用RosetteSep™人造血祖细胞富集抗体混合物 (产品号 #15186C)预富集造血祖细胞，包含可识别CD2, CD3, CD14, CD16, CD19, CD24, CD56, CD66b和CD61细胞表面标志物的抗体。然后使用EasySep™人CD34正选抗体混合物 (产品号 #18066C) 来分选CD34+细胞，其中包含识别CD34的抗体。RosetteSep™将非目的细胞与红细胞 (RBCs) 交联，形成免疫玫瑰花环，并在密度梯度离心时沉降。含CD34+细胞的预富集组分可从血浆和密度梯度离心液介质的界面收集。预富集的CD34+细胞随后用抗体和磁珠标记，并通过EasySep™磁极，无需分离柱分选。非目的细胞通过简单倾倒弃去，而目的细胞则保留在管中。分选后的CD34+细胞可立即用于下游应用。

首先，使用RosetteSep™人造血祖细胞富集抗体混合物 (产品号 #15186C)预富集造血祖细胞，包含可识别CD2, CD3, CD14, CD16, CD19, CD24, CD56, CD66b和CD61细胞表面标志物的抗体。然后使用EasySep™人CD34正选抗体混合物 (产品号 #18066C) 来分选CD34+细胞，其中包含识别CD34的抗体。RosetteSep™将非目的细胞与红细胞 (RBCs) 交联，形成免疫玫瑰花环，并在密度梯度离心时沉降。含CD34+细胞的预富集组分可从血浆和密度梯度离心液介质的界面收集。预富集的CD34+细胞随后用抗体和磁珠标记，并通过EasySep™磁极，无需分离柱分选。非目的细胞通过简单倾倒弃去，而目的细胞则保留在管中。分选后的CD34+细胞可立即用于下游应用。

磁体兼容性
• “The Big Easy” EasySep™ Magnet (Catalog #18001) • RoboSep™-S (Catalog #21000)
 
亚型
细胞分选试剂盒
 
细胞类型
造血干/祖细胞
 
种属
人
 
样本来源
Buffy Coat，Whole Blood
 
筛选方法
Positive
 
应用
细胞分选
 
品牌
EasySep，RoboSep，RosetteSep
 
研究领域
免疫，干细胞生物学
 

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实验数据

Figure 1.Typical CD34 Selection Profile on Human Whole Blood

Starting with whole peripheral blood, the CD34+ cell content of the isolated fraction is typically 95.1 ± 4.5% (gated on viable CD45+ cells; mean ± SD for the silver “The Big Easy” EasySep™ Magnet). In the above example, the purities of the start and final isolated fractions are 0.06% and 96.2%, respectively.

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
Complete Kit for Human Whole Blood CD34+ Cells
Catalog #
15086
Lot #
All
Language
English
Document Type
Product Information Sheet
Product Name
RoboSep™ Complete Kit for Human Whole Blood CD34+ Cells
Catalog #
15086RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Product Name
Complete Kit for Human Whole Blood CD34+ Cells
Catalog #
15086
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Product Name
Complete Kit for Human Whole Blood CD34+ Cells
Catalog #
15086
Lot #
All
Language
English
Document Type
Safety Data Sheet 3
Product Name
Complete Kit for Human Whole Blood CD34+ Cells
Catalog #
15086
Lot #
All
Language
English
Document Type
Safety Data Sheet 4
Product Name
Complete Kit for Human Whole Blood CD34+ Cells
Catalog #
15086
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Product Name
RoboSep™ Complete Kit for Human Whole Blood CD34+ Cells
Catalog #
15086RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Product Name
RoboSep™ Complete Kit for Human Whole Blood CD34+ Cells
Catalog #
15086RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 3
Product Name
RoboSep™ Complete Kit for Human Whole Blood CD34+ Cells
Catalog #
15086RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 4
Product Name
RoboSep™ Complete Kit for Human Whole Blood CD34+ Cells
Catalog #
15086RF
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 (5)

Human CD34+ Cell Isolation Product Selection
Brochure
Human CD34+ Cell Isolation Product Selection
Hematopoietic Stem and Progenitor Cells - Products for Your Research
Brochure
Hematopoietic Stem and Progenitor Cells - Products for Your Research
Frequencies of Human Cell Types in Blood-Related Sources
Wallchart
Frequencies of Human Cell Types in Blood-Related Sources
Human Immune Cytokines
Wallchart
Human Immune Cytokines
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

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 (1)

Nucleosome Density ChIP-Seq Identifies Distinct Chromatin Modification Signatures Associated with MNase Accessibility. Lorzadeh A et al. Cell reports 2016 NOV

Abstract

Nucleosome position, density, and post-translational modification are widely accepted components of mechanisms regulating DNA transcription but still incompletely understood. We present a modified native ChIP-seq method combined with an analytical framework that allows MNase accessibility to be integrated with histone modification profiles. Application of this methodology to the primitive (CD34+) subset of normal human cord blood cells enabled genomic regions enriched in one versus two nucleosomes marked by histone 3 lysine 4 trimethylation (H3K4me3) and/or histone 3 lysine 27 trimethylation (H3K27me3) to be associated with their transcriptional and DNA methylation states. From this analysis, we defined four classes of promoter-specific profiles and demonstrated that a majority of bivalent marked promoters are heterogeneously marked at a single-cell level in this primitive cell type. Interestingly, extension of this approach to human embryonic stem cells revealed an altered relationship between chromatin modification state and nucleosome content at promoters, suggesting developmental stage-specific organization of histone methylation states.
View publication
View All Publications

更多信息

更多信息
种属 Human
Magnet Compatibility • “The Big Easy” EasySep™ Magnet (Catalog #18001) • RoboSep™-S (Catalog #21000)
样本来源 Buffy Coat, Whole Blood
Selection Method Positive
标记抗体
  1. 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

  2. 抗人CD45抗体，克隆HI30
    抗人CD45抗体，克隆HI30

    小鼠单克隆IgG1抗体，抗人、黑猩猩CD45

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