EasySep™小鼠造血祖细胞分选试剂盒

通过免疫磁珠负选分离出无磁珠标记的小鼠干细胞与祖细胞

产品号 #(选择产品)

产品号 #19856_C

通过免疫磁珠负选分离出无磁珠标记的小鼠干细胞与祖细胞

产品优势

  • 易于操作、快速
  • 纯度高达84%
  • 无需分离柱

产品组分包括

  • EasySep™小鼠造血祖细胞分选试剂盒(产品号 #19856)
    • EasySep™小鼠造血祖细胞分选抗体混合物,0.5 mL
    • EasySep™ Streptavidin RapidSpheres™ 50001磁珠,1 mL
    • EasySep™ 小鼠FcR阻断剂(产品号#18730),0.2 mL
  • RoboSep™小鼠造血祖细胞分选试剂盒产品号 #19856RF)
    • EasySep™小鼠造血祖细胞分选抗体混合物,0.5 mL
    • EasySep™ Streptavidin RapidSpheres™ 50001磁珠,1 mL
    • EasySep™ 小鼠FcR阻断剂(产品号#18730),0.2 mL
    • RoboSep™ 缓冲液(产品号#20104)
    • RoboSep™过滤吸头(产品号#20125)
New format, same high quality! You may notice that your kit contents and packaging look slightly different from previous orders. We are currently updating the format of select EasySep™ Mouse kits to include a Mouse FcR blocker instead of Normal Rat Serum. With this change, all components will now be shipped in a single package, while providing the same cell isolation performance as before.

总览

使用EasySep™小鼠造血祖细胞分选试剂盒,通过免疫磁珠负选技术,可轻松高效地从骨髓样本中分离高纯度小鼠祖细胞。当使用其他类型组织来源的单细胞悬液时,该试剂盒可能需要优化。

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。

在此 EasySep™ 负选方案中,非目的细胞通过抗体复合物与磁珠标记。非目的细胞可通过针对非造血干细胞和非祖细胞(CD5、CD11b、CD19、CD45R/B220、Ly6G/C(Gr-1)、TER119、7-4)的抗体而被去除。通过使用 EasySep™ 磁力架,将被磁珠标记的非目的细胞分离,仅需简单倒出,即可获得无磁珠标记的目标小鼠造血细胞。完成磁珠分选后,目标小鼠造血细胞可直接用于流式细胞术、细胞培养等下游实验。

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。探索为您的实验流程优化的更多产品,包括培养基、添加剂、抗体等。

磁体兼容性
• EasySep™ Magnet (Catalog #18000) • “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
小鼠
 
样本来源
Bone Marrow
 
筛选方法
Negative
 
应用
细胞分选
 
品牌
EasySep,RoboSep
 
研究领域
免疫,干细胞生物学
 

Data Figures

Typical EasySep™ Mouse Hematopoietic Progenitor Cell Isolation Profile

Figure 1. Typical EasySep™ Mouse Hematopoietic Progenitor Cell Isolation Profile

Starting with a mouse bone marrow cell suspension, the lineage antigen-negative cell content of the isolated fraction typically ranges from 60 - 84%.

Protocols and Documentation

Find supporting information and directions for use in the Product Information Sheet or explore additional protocols below.

Document Type
Product Name
Catalog #
Lot #
Language
Catalog #
19856RF
Lot #
1000138480 or lower
Language
English
Catalog #
19856RF
Lot #
1000138481 or higher
Language
English
Catalog #
19856
Lot #
1000138481 or higher
Language
English
Catalog #
19856
Lot #
1000138480 or lower
Language
English
Document Type
Safety Data Sheet 1
Catalog #
19856RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Catalog #
19856RF
Lot #
All
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English
Document Type
Safety Data Sheet 3
Catalog #
19856RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 4
Catalog #
19856RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 5
Catalog #
19856RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Catalog #
19856
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Catalog #
19856
Lot #
All
Language
English
Document Type
Safety Data Sheet 3
Catalog #
19856
Lot #
All
Language
English
Document Type
Safety Data Sheet 4
Catalog #
19856
Lot #
All
Language
English

Applications

This product is designed for use in the following research area(s) as part of the highlighted workflow stage(s). Explore these workflows to learn more about the other products we offer to support each research area.

Resources and Publications

Educational Materials (4)

Frequently Asked Questions

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.

Publications (9)

Intra-Peritoneal Transplantation for Generating Acute Myeloid Leukemia in Mice. F. Qian et al. Journal of visualized experiments : JoVE 2023 jan

Abstract

There is an unmet need for novel therapies to treat acute myeloid leukemia (AML) and the associated relapse that involves persistent leukemia stem cells (LSCs). An experimental AML rodent model to test therapies based on successfully transplanting these cells via retro-orbital injections in recipient mice is fraught with challenges. The aim of this study was to develop an easy, reliable, and consistent method to generate a robust murine model of AML using an intra-peritoneal route. In the present protocol, bone marrow cells were transduced with a retrovirus expressing human MLL-AF9 fusion oncoprotein. The efficiency of lineage negative (Lin-) and Lin-Sca-1+c-Kit+ (LSK) populations as donor LSCs in the development of primary AML was tested, and intra-peritoneal injection was adopted as a new method to generate AML. Comparison between intra-peritoneal and retro-orbital injections was done in serial transplantations to compare and contrast the two methods. Both Lin- and LSK cells transduced with human MLL-AF9 virus engrafted well in the bone marrow and spleen of recipients, leading to a full-blown AML. The intra-peritoneal injection of donor cells established AML in recipients upon serial transplantation, and the infiltration of AML cells was detected in the blood, bone marrow, spleen, and liver of recipients by flow cytometry, qPCR, and histological analyses. Thus, intra-peritoneal injection is an efficient method of AML induction using serial transplantation of donor leukemic cells.
Interleukin-4 treatment reduces leukemia burden in acute myeloid leukemia. F. Qian et al. FASEB journal : official publication of the Federation of American Societies for Experimental Biology 2022 may

Abstract

Interleukin-4 (IL-4) is a signature cytokine pivotal in Type 2 helper T cell (Th2) immune response, particularly in allergy and hypersensitivity. Interestingly, IL-4 increases endogenous levels of prostaglandin D2 (PGD2 ) and its metabolites, $\Delta$12 -prostaglandin J2 ($\Delta$12 -PGJ2 ) and 15-deoxy-$\Delta$12,14 -prostaglandin J2 (15d-PGJ2 ), collectively called cyclopentenone PGs (CyPGs). However, the therapeutic role of IL-4 in hematologic malignancies remains unclear. Here, we employed a murine model of acute myeloid leukemia (AML), where human MLL-AF9 fusion oncoprotein was expressed in hematopoietic progenitor cells, to test the effect of IL-4 treatment in vivo. Daily intraperitoneal treatment with IL-4 at 60 µg/kg/d significantly alleviated the severity of AML, as seen by decreased leukemia-initiating cells (LICs). The effect of IL-4 was mediated, in part, by the enhanced expression of hematopoietic- PGD2  synthase (H-PGDS) to effect endogenous production of CyPGs, through autocrine and paracrine signaling mechanisms. Similar results were seen with patient-derived AML cells cultured ex vivo with IL-4. Use of GW9662, a peroxisome proliferator-activated receptor gamma (PPAR$\gamma$) antagonist, suggested endogenous CyPGs-PPAR$\gamma$ axis mediated p53-dependent apoptosis of LICs by IL-4. Taken together, our results reveal a beneficial role of IL-4 treatment in AML suggesting a potential therapeutic regimen worthy of clinical trials in patients with AML.
Protocol to identify and analyze mouse and human quiescent hematopoietic stem cells using flow cytometry combined with confocal imaging. J. Qiu et al. STAR protocols 2022 dec

Abstract

Mitochondrial membrane potential (MMP) segregates functionally distinct subsets within highly purified hematopoietic stem cells (HSCs). Here, we detail a protocol for FACS isolation of MMP sub-fractions of phenotypically defined mouse and human HSCs. These steps are followed by high-/super-resolution immunofluorescence microscopy of HSCs' lysosomes. While the protocol describes the isolation of quiescent HSCs, which are the most potent subsets, it could also be applied to other HSC subsets. This protocol overcomes some experimental challenges associated with low HSC numbers. For complete details on the use and execution of this protocol, please refer to Liang et al. (2020) and Qiu et al. (2021).

更多信息

更多信息
Species Mouse
Magnet Compatibility • EasySep™ Magnet (Catalog #18000) • “The Big Easy” EasySep™ Magnet (Catalog #18001) • RoboSep™-S (Catalog #21000)
Sample Source Bone Marrow
Selection Method Negative
标记抗体
PRODUCTS ARE FOR RESEARCH USE ONLY AND NOT INTENDED FOR HUMAN OR ANIMAL DIAGNOSTIC OR THERAPEUTIC USES UNLESS OTHERWISE STATED. FOR ADDITIONAL INFORMATION ON QUALITY AT STEMCELL, REFER TO WWW.STEMCELL.COM/COMPLIANCE.
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