EasySep™ Direct Human Naïve B Cell Isolation Kit

Immunomagnetic negative selection kit from whole blood
EasySep™ Direct Human Naïve B Cell Isolation Kit

Immunomagnetic isolation of naïve B cells directly from whole blood

100 mL whole blood
Catalog # 19264
545 USD
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Required Products
  1. EasySep™ Magnet
    EasySep™ Magnet

    Magnet for column-free immunomagnetic separation

Overview

This kit is used to isolate highly purified naïve B cells directly from human whole blood. Isolated cells are unlabeled with antibodies or particles and undergo minimal manipulation.

This kit targets non-naïve B cells for removal with antibodies recognizing specific surface markers. Unwanted cells are labeled with antibodies and EasySep™ Direct RapidSpheres™, and separated using an EasySep™ magnet. Desired cells are simply collected into a new tube and are immediately available for downstream applications such as flow cytometry, culture or DNA/RNA extraction.
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
Components
  • EasySep™ Direct Human Naïve B Cell Isolation Kit (Catalog #19264)
    • EasySep™ Direct Human Naïve B Cell Isolation Cocktail, 2 x 2.5 mL
    • EasySep™ Direct RapidSpheres™, 4 x 2.5 mL
Magnet Compatibility
• EasySep™ Magnet (Catalog #18000)
• “The Big Easy” EasySep™ Magnet (Catalog #18001)
• Easy 50 EasySep™ Magnet (Catalog #18002)
• EasyEights™ EasySep™ Magnet (Catalog #18103)
Subtype
Cell Isolation Kits
Cell Type
B Cells
Species
Human
Sample Source
Whole Blood
Selection Method
Negative
Application
Cell Isolation
Brand
EasySep
Area of Interest
Immunology

Scientific Resources

Product Documentation

Document Type Product Name Catalog # Lot # Language
Document Type
Product Information Sheet
Product Name
EasySep™ Direct Human Naïve B Cell Isolation Kit
Catalog #
19264
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Product Name
EasySep™ Direct Human Naïve B Cell Isolation Kit
Catalog #
19264
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Product Name
EasySep™ Direct Human Naïve B Cell Isolation Kit
Catalog #
19264
Lot #
All
Language
English

Educational Materials (12)

Brochure
Tools For Your Immunology Research
Brochure
EasySep™ Cell Separation Technology
Brochure
Human B Cell Isolation Product Selection
Brochure
EasySep™ Direct: Cell Isolation from Whole Blood in 20 Minutes
Technical Bulletin
High-Throughput Immunodensity-Based Cell Subset Isolation Directly from Whole Blood
Wallchart
Frequencies of Cell Types in Human Peripheral Blood
Wallchart
Human Immune Cytokines
Video
Cell Isolation Directly from Whole Blood without RBC Lysis or Centrifugation: EasySep™ Direct
1:44
Cell Isolation Directly from Whole Blood without RBC Lysis or Centrifugation: EasySep™ Direct
Video
How to Isolate Cells Directly from Whole Blood Using the Easy 50 EasySep™ Magnet: 30-Minute Protocol
5:17
How to Isolate Cells Directly from Whole Blood Using the Easy 50 EasySep™ Magnet: 30-Minute Protocol
Video
How to Isolate Cells Directly from Whole Blood - EasySep™ Direct Protocol, EasyEights Magnet
6:13
How to Isolate Cells Directly from Whole Blood - EasySep™ Direct Protocol, EasyEights Magnet
Video
How to Isolate Cells Directly from Whole Blood Using the EasySep™ Purple/Silver Magnets
4:30
How to Isolate Cells Directly from Whole Blood Using the EasySep™ Purple/Silver Magnets
Video
Isolate Cells with a Simple Pour-Off: EasySep™ Cell Separation Technology
1:13
Isolate Cells with a Simple Pour-Off: EasySep™ Cell Separation Technology

Frequently Asked Question

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.
Read More

Product 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.

Data and Publications

Data

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).

Publications (1)

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

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

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