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EasySep™人脐带血CD34正选试剂盒II

脐带血中人CD34+细胞的免疫磁珠正选
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产品号 #（选择产品）

产品号 #17896_C

脐带血中人CD34+细胞的免疫磁珠正选

产品优势

  • 快捷、操作简单
  • 纯度可高达98%
  • 无需分离柱

产品组分包括

  • EasySep™人脐带血CD34+正选试剂盒II (产品号 #17896)
    • RosetteSep™脐带血CD34预富集抗体混合物II，2x2.5mL
    • EasySep™人CD34正选抗体混合物，2x1mL
    • EasySep™ Dextran RapidSpheres™ 50100 磁珠，2x1mL
    • RoboSep™人脐带血CD34正选试剂盒II    
    • RosetteSep™脐带血CD34预富集抗体混合物II，2x2.5mL
    • EasySep™人CD34正选抗体混合物，2x1mL
    • EasySep™ Dextran RapidSpheres™ 50100 磁珠，2x1mL
    • RoboSep™ 缓冲液（产品号 #20104）
    • RoboSep™过滤吸头（产品号#20125）x 2
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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™
  2. EasySep™磁极
    EasySep™磁极
  3. EasySep™缓冲液
    EasySep™缓冲液
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™人脐带血CD34正选试剂盒II，通过免疫磁珠正选技术，从新鲜脐带血全血样本中分离高纯度人CD34+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。

首先，使用RosetteSep™人脐带血CD34预富集抗体混合物 (产品号 #15896C) 识别T细胞、B细胞、髓系细胞以及血小板表面标志物的抗体来预富集造血祖细胞。然后使用EasySep™人CD34正选抗体混合物 (产品号 #18096C) 中识别CD34的抗体来分选CD34+细胞。抗体复合物中还含有人Fc受体抗体，可最大程度地减少非特异性结合。使用EasySep™磁力架分离标记细胞，只需简单倾倒或枪头吸取非目标细胞，目标细胞保留在管中。经磁珠分选后，目标CD34+细胞即可直接用于下游实验。

从新鲜的人外周血或白膜分离CD34+细胞，推荐使用人全血CD34+细胞全套试剂盒（产品号 #15086）。

如需从其他样本分离CD34+细胞，包括新鲜或冻存的动员外周血、骨髓单个核细胞、冻存的脐带血单个核细胞等，推荐 EasySep™人CD34正选试剂盒II（产品号 #17856）。

本产品替代独立包装的EasySep™人脐带血CD34正选试剂盒（产品号#18096）。

了解更多关于免疫磁珠EasySep™技术的工作原理，或如何通过RoboSep™实现免疫磁珠细胞分选全自动化，探索更多产品优化您的实验流程，包括培养基、添加剂、抗体等。

 

磁极兼容性
• EasySep™磁极（产品号 #18000）
• “The Big Easy” EasySep™磁极（产品号 #18001）
• EasyEights™ EasySep™磁极（产品号 #18103）
• RoboSep™-S（产品号 #21000）
 
分类
细胞分选试剂盒
 
细胞类型
造血干/祖细胞
 
种属
人
 
样本来源
脐带血
 
分选方法
正选
 
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

Typical EasySep™ Human Cord Blood CD34 Positive Selection Profile

Figure 1. Typical EasySep™ Human Cord Blood CD34 Positive Selection Profile

Starting with fresh cord blood, the CD34+ cell content of the isolated fraction is typically 91 ± 9% (mean ± SD) using the purple EasySep™ Magnet.

Isolation of CD34+ Cells Using EasySep™ Human Cord Blood CD34 Positive Selection Kit II

Figure 2. Isolation of CD34+ Cells Using EasySep™ Human Cord Blood CD34 Positive Selection Kit II

CD45 and CD34 expression of cells before separation (“Start”), after RosetteSep™ (“Pre-Enriched”), and after selection of CD34+ cells (“Isolated”) using EasySep™ Human Cord Blood CD34 Positive Selection Kit II (Catalog #17896). Results of a typical experiment are shown. CD45+CD34+ HSPCs (top right quadrant) have been enriched > 15-fold (from 0.6% to 10%) after RosetteSep™ pre-enrichment and > 200-fold (from 0.6% to 98%) after EasySep™ CD34+ selection (“Isolated”). The flow cytometry data shown are gated on cells with intermediate-to-high forward light scatter (FSC) that are negative for propidium iodide (PI) staining to exclude debris, RBCs, platelets, and dead cells. Based on the results of cell separations with 15 different CB samples with a starting CD34+ cell purity of 0.4%, the average CD34+ cell purity is 6% after RosetteSep™ pre-enrichment and 91% after EasySep™ cell isolation.

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
产品说明书
Product Name
RoboSep™ 人脐带血CD34正选试剂盒II
Catalog #
17896RF
Lot #
All
Language
中文
Document Type
产品说明书
Product Name
EasySep™人脐带血CD34正选试剂盒II
Catalog #
17896
Lot #
All
Language
中文
Document Type
Product Information Sheet
Product Name
RoboSep™ Human CB CD34 Positive Selection Kit II
Catalog #
17896RF
Lot #
All
Language
English
Document Type
Product Information Sheet
Product Name
EasySep™ Human Cord Blood CD34 Positive Selection Kit II
Catalog #
17896
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Product Name
RoboSep™ Human CB CD34 Positive Selection Kit II
Catalog #
17896RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Product Name
RoboSep™ Human CB CD34 Positive Selection Kit II
Catalog #
17896RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 3
Product Name
RoboSep™ Human CB CD34 Positive Selection Kit II
Catalog #
17896RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 4
Product Name
RoboSep™ Human CB CD34 Positive Selection Kit II
Catalog #
17896RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Product Name
EasySep™ Human Cord Blood CD34 Positive Selection Kit II
Catalog #
17896
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Product Name
EasySep™ Human Cord Blood CD34 Positive Selection Kit II
Catalog #
17896
Lot #
All
Language
English
Document Type
Safety Data Sheet 3
Product Name
EasySep™ Human Cord Blood CD34 Positive Selection Kit II
Catalog #
17896
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 (10)

Human CD34+ Cell Isolation Product Selection
产品手册
Human CD34+ Cell Isolation Product Selection
EasySep™细胞分选技术 - 更高效地分离细胞
产品手册
EasySep™细胞分选技术 - 更高效地分离细胞
Your Ideas. Our Tools. 为您提供造血干细胞和祖细胞研究 各个步骤的相关产品
产品手册
Your Ideas. Our Tools. 为您提供造血干细胞和祖细胞研究 各个步骤的相关产品
从人脐带血分离CD34+细胞
技术公告
从人脐带血分离CD34+细胞
Generation of B Cells from Cord Blood-Derived CD34+ Cells Using StemSpan™ Medium and Supplements
技术公告
Generation of B Cells from Cord Blood-Derived CD34+ Cells Using StemSpan™ Medium and Supplements
Isolate Cells with a Simple Pour-Off: EasySep™ Cell Separation Technology
视频
Isolate Cells with a Simple Pour-Off: EasySep™ Cell Separation Technology
How to Isolate PBMCs from Whole Blood Using Density Gradient Centrifugation (Ficoll™ or Lymphoprep™)
视频
How to Isolate PBMCs from Whole Blood Using Density Gradient Centrifugation (Ficoll™ or Lymphoprep™)
Simultaneous Cell Isolation from Multiple Samples Using the EasyEights™ EasySep™ Magnet
视频
Simultaneous Cell Isolation from Multiple Samples Using the EasyEights™ EasySep™ Magnet
Troubleshooting In Vitro Expansion of Leukemic Cells
视频
Troubleshooting In Vitro Expansion of Leukemic Cells
Human Immune Cytokines
挂图
Human Immune Cytokines

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 (9)

Directed evolution of a recombinase that excises the provirus of most HIV-1 primary isolates with high specificity. Karpinski J et al. Nature Biotechnology 2016 APR

Abstract

Current combination antiretroviral therapies (cART) efficiently suppress HIV-1 reproduction in humans,but the virus persists as integrated proviral reservoirs in small numbers of cells. To generate an antiviral agent capable of eradicating the provirus from infected cells,we employed 145 cycles of substrate-linked directed evolution to evolve a recombinase (Brec1) that site-specifically recognizes a 34-bp sequence present in the long terminal repeats (LTRs) of the majority of the clinically relevant HIV-1 strains and subtypes. Brec1 efficiently,precisely and safely removes the integrated provirus from infected cells and is efficacious on clinical HIV-1 isolates in vitro and in vivo,including in mice humanized with patient-derived cells. Our data suggest that Brec1 has potential for clinical application as a curative HIV-1 therapy.
View publication
Discovery of Berberine that Targetedly Induces Autophagic Degradation of both BCR-ABL and BCR-ABL T315I through Recruiting LRSAM1 for Overcoming Imatinib Resistance. Z. Yin et al. Clinical cancer research : an official journal of the American Association for Cancer Research 2020 feb

Abstract

PURPOSE Imatinib,the breakpoint cluster region protein (BCR)/Abelson murine leukemia viral oncogene homolog (ABL) inhibitor,is widely used to treat chronic myeloid leukemia (CML). However,imatinib resistance develops in many patients. Therefore,new drugs with improved therapeutic effects are urgently needed. Berberine (BBR) is a potent BCR-ABL inhibitor for imatinib-sensitive and -resistant CML. EXPERIMENTAL DESIGN Protein structure analysis and virtual screening were used to identify BBR targets in CML. Molecular docking analysis,surface plasmon resonance imaging,nuclear magnetic resonance assays,and thermoshift assays were performed to confirm the BBR target. The change in BCR-ABL protein expression after BBR treatment was assessed by Western blotting. The effects of BBR were assessed in vitro in cell lines,in vivo in mice,and in human CML bone marrow cells as a potential strategy to overcome imatinib resistance. RESULTS We discovered that BBR bound to the protein tyrosine kinase domain of BCR-ABL. BBR inhibited the activity of BCR-ABL and BCR-ABL with the T315I mutation,and it also degraded these proteins via the autophagic lysosome pathway by recruiting E3 ubiquitin-protein ligase LRSAM1. BBR inhibited the cell viability and colony formation of CML cells and prolonged survival in CML mouse models with imatinib sensitivity and resistance. CONCLUSIONS The results show that BBR directly binds to and degrades BCR-ABL and BCR-ABL T315I via the autophagic lysosome pathway by recruiting LRSAM1. The use of BBR is a new strategy to improve the treatment of patients with CML with imatinib sensitivity or resistance.
View publication
Reducing TGF-$\beta$1 cooperated with StemRegenin 1 promoted the expansion ex vivo of cord blood CD34+ cells by inhibiting AhR signalling. X. Zhu et al. Cell proliferation 2021 mar

Abstract

OBJECTIVE As an inhibitor of the AhR signalling pathway,StemRegenin 1 (SR1) not only promotes the expansion of CD34+ cells but also increases CD34- cell numbers. These CD34- cells influenced the ex vivo expansion of CD34+ cells. In this work,the effects of periodically removing CD34- cells combined with SR1 addition on the ex vivo expansion and biological functions of HSCs were investigated. MATERIALS AND METHODS CD34- cells were removed periodically with SR1 addition to investigate cell subpopulations,cell expansion,biological functions,expanded cell division mode and supernatant TGF-$\beta$1 contents. RESULTS After 10-day culture,the expansion of CD34+ cells in the CD34- cell removal plus SR1 group was significantly higher than that in the control group and the SR1 group. Moreover,periodically removing CD34- cells with SR1 addition improved the biological function of expanded CD34+ cells and significantly increased the percentage of self-renewal symmetric division of CD34+ cells. In addition,the concentration of total TGF-$\beta$1 and activated TGF-$\beta$1 in the supernatant was significantly lower than those in the control group and the SR1 group. RT-qPCR results showed that the periodic removal of CD34- cells with cooperation from SR1 further reduced the expression of AhR-related genes. CONCLUSIONS Periodic removal of CD34- cells plus cooperation with SR1 improved the expansion of CD34+ cells,maintained better biological function of expanded CD34+ cells and reduced the TGF-$\beta$1 contents by downregulating AhR signalling.
View publication
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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) • EasyEights™ EasySep™ Magnet (Catalog #18103) • RoboSep™-S (Catalog #21000)
样本来源 脐带血
Selection Method Positive
标记抗体
  1. 抗人CD34抗体，clone 8G12
    抗人CD34抗体，clone 8G12

    小鼠Monoclonal IgG1抗体，抗人CD34

  2. 抗人CD45抗体，clone HI30
    抗人CD45抗体，clone HI30

    小鼠Monoclonal IgG1抗体，抗人、黑猩猩CD45

  3. 抗人CD34抗体，clone 581
    抗人CD34抗体，clone 581

    小鼠Monoclonal IgG1抗体，抗人CD34

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