EasySep? Direct Human Na?ve B Cell Isolation Kit

Immunomagnetic isolation of na?ve B cells directly from whole blood

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EasySep? Direct Human Na?ve B Cell Isolation Kit

Immunomagnetic isolation of na?ve B cells directly from whole blood

Size

100 mL whole blood processed Request Pricing

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Immunomagnetic isolation of na?ve B cells directly from whole blood

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Product Advantages

> 99.9% red blood cell (RBC) depletion without the need for density gradient centrifugation, sedimentation, or lysis

Fast, easy-to-use and column-free

Up to 95% purity of isolated cells

Isolated cells are untouched

What's Included

EasySep? Direct Human Na?ve B Cell Isolation Kit (Catalog #19264)

EasySep? Direct Human Na?ve B Cell Isolation Cocktail, 2 x 2.5 mL
EasySep? Direct RapidSpheres?, 4 x 2.5 mL

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Overview

Easily and efficiently isolate highly purified human na?ve B cells directly from human whole blood samples by immunomagnetic negative selection, with the EasySep? Direct Human Na?ve B Cell Isolation Kit. Widely used in published research for more than 20 years, EasySep? combines the specificity of monoclonal antibodies with the simplicity of a column-free magnetic system.

In this EasySep? negative selection procedure, unwanted cells are labeled with antibody complexes and magnetic particles called EasySep? Direct RapidSpheres?. Unwanted cells expressing the following markers are targeted for removal: CD2, CD3, CD15, CD16, CD27, CD36, CD43, CD56, CD66b, and GlyA. The magnetically labeled cells are then separated from the untouched desired na?ve B cells by using an EasySep? magnet and simply pouring or pipetting the desired cells into a new tube. Following magnetic cell isolation, the desired na?ve B cells are ready for downstream applications such as flow cytometry, culture, or DNA/RNA extraction.

Learn more about how immunomagnetic EasySep? technology works. Explore additional products optimized for your workflow, including those for cell characterization, cryopreservation, and more.

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Cell Isolation Kits

B Cells

Human

Whole Blood

Negative

Cell Isolation

EasySep

Immunology

Data Figures

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

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).

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

Document Type

Product Name

Catalog #

19264

Lot #

All

Language

English

Document Type

Product Name

Catalog #

19264

Lot #

All

Language

English

Document Type

Product Name

Catalog #

19264

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.

Research Area

Workflow Stages

Workflow Stages for B Cell Research

Cell Sourcing

Cell Isolation

Cell Expansion

Cell Characterization

Resources and Publications

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.

BCR-associated factors driving chronic lymphocytic leukemia cells proliferation ex vivo. C. Schleiss et al. Scientific reports 2019 jan

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.

View All Publications

Labeling Antibodies

Anti-Human CD19 Antibody, Clone HIB19
Mouse monoclonal IgG1 antibody against human, chimpanzee CD19
Anti-Human CD45 Antibody, Clone HI30
Mouse monoclonal IgG1 antibody against human, chimpanzee CD45

Quality Statement:

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 ��ƽ��, REFER TO WWW.��ƽ��.COM/COMPLIANCE.

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EasySep? Direct Human Na?ve B Cell Isolation Kit

Immunomagnetic isolation of na?ve B cells directly from whole blood

EasySep? Direct Human Na?ve B Cell Isolation Kit

Immunomagnetic isolation of na?ve B cells directly from whole blood

Product Advantages

What's Included

Request Pricing

Overview

Data Figures

Protocols and Documentation

Applications

Resources and Publications

Educational Materials (13)

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

Related Products

Labeling Antibodies

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