EasySep™ Direct人Naïve B细胞分选试剂盒

全血免疫磁珠负选试剂盒

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产品号 #19264

直接从全血中通过免疫磁珠分选naïve B细胞

产品优势

> 99.9%的红细胞（RBC）去除率，无需密度梯度离心、沉降或裂解
易于操作、快速，且无需分离柱
分选获得的细胞纯度高达95%
分选得到的细胞未被标记

产品组分包括

EasySep™ Direct人Naïve B细胞分选试剂盒（产品号#19264）
- EasySep™ Direct人naïve B细胞分选抗体混合物，2 x 2.5 mL
- EasySep™ Direct RapidSpheres™磁珠，4 x 2.5 m

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EasySep™ Direct人Naïve B细胞分选试剂盒

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实验数据

$Starting with human whole blood from normal healthy donors, the typical naïve B cell (CD19+CD27-) content of the non-lysed final isolated fraction is 918 ± 36% (gated on CD45) or 824 ± 126% (not gated on CD45)$

Figure 1. Typical EasySep™ Direct Human Naïve B Cell Isolation Profile

Starting with human whole blood from normal healthy donors, the typical naïve B cell (CD19+CD27-) content of the non-lysed final isolated fraction is 91.8 ± 3.6% (gated on CD45) or 82.4 ± 12.6% (not gated on CD45). In the above example, the naïve B cell (CD19+CD27-) content of the lysed whole blood start sample and non-lysed final isolated fraction is 5.1% and 91.5% (gated on CD45), respectively, or 5.1% and 90.0% (not gated on CD45), respectively. The starting frequency of naïve B cells in the non-lysed whole blood start sample above is 0.009% (data not shown).

产品说明书及文档

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

应用领域

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.

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 (1)

Abstract

A chronic antigenic stimulation is believed to sustain the leukemogenic development of chronic lymphocytic leukemia (CLL) and most of lymphoproliferative malignancies developed from mature B cells. Reproducing a proliferative stimulation ex vivo is critical to decipher the mechanisms of leukemogenesis in these malignancies. However, functional studies of CLL cells remains limited since current ex vivo B cell receptor (BCR) stimulation protocols are not sufficient to induce the proliferation of these cells, pointing out the need of mandatory BCR co-factors in this process. Here, we investigated benefits of several BCR co-stimulatory molecules (IL-2, IL-4, IL-15, IL-21 and CD40 ligand) in multiple culture conditions. Our results demonstrated that BCR engagement (anti-IgM ligation) concomitant to CD40 ligand, IL-4 and IL-21 stimulation allowed CLL cells proliferation ex vivo. In addition, we established a proliferative advantage for ZAP70 positive CLL cells, associated to an increased phosphorylation of ZAP70/SYK and STAT6. Moreover, the use of a tri-dimensional matrix of methylcellulose and the addition of TLR9 agonists further increased this proliferative response. This ex vivo model of BCR stimulation with T-derived cytokines is a relevant and efficient model for functional studies of CLL as well as lymphoproliferative malignancies.

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EasySep™ Direct人Naïve B细胞分选试剂盒

产品号 #19264

产品优势

产品组分包括

概述

实验数据

产品说明书及文档

应用领域

相关材料与文献

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Scientists Helping Scientists™

EasySep™ Direct人Naïve B细胞分选试剂盒

产品号 #19264

产品优势

产品组分包括

概述

实验数据

产品说明书及文档

应用领域

相关材料与文献

Educational Materials (12)

Frequently Asked Questions

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?

Publications (1)

Abstract

Request Pricing

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