EasySep™ Direct Human B-CLL Cell Isolation Kit

Immunomagnetic negative selection from whole blood kit

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EasySep™ Direct Human B-CLL Cell Isolation Kit

Immunomagnetic isolation of B-CLL cells directly from whole blood

100 mL whole blood
Catalog #19664
565 USD

Required Products

Overview

This kit isolates untouched and highly purified B-CLL cells directly from human whole blood by fully immunomagnetic negative selection. The depletion cocktail in this kit does not contain antibodies against CD43, which can be expressed on malignant cells in chronic lymphocytic leukemia (CLL) samples.

This cell isolation kit targets non-B cells for removal based on specific surface antigens. 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 can immediately be used in downstream applications such as nucleic acid extraction, flow cytometry or culture.
Advantages:
• > 99.9% RBC depletion without the need for density gradient centrifugation, sedimentation, or lysis
• Fast, easy-to-use and column-free
• Isolated cells are untouched
Components:
  • EasySep™ Direct Human B-CLL Cell Isolation Kit (Catalog #19664)
    • EasySep™ Direct Human B-CLL 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; Cancer Research

Scientific Resources

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.
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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.

Research Area Workflow Stages for
Workflow Stages

Data and Publications

Data

Starting with human whole blood from normal healthy donors, the typical B cell (CD3-CD19+) content of the non-lysed final isolated fraction is 87.0 ± 7.6% (gated on CD45) or 84.1 ± 11.7% (not gated on CD45)

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

Starting with human whole blood from normal healthy donors, the typical B cell (CD3-CD19+) content of the non-lysed final isolated fraction is 87.0 ± 7.6% (gated on CD45) or 84.1 ± 11.7% (not gated on CD45). In the example above, the B cell (CD3-CD19+) content of the lysed whole blood start and non-lysed final isolated fraction is 4.3% and 88.0% (gated on CD45), respectively, or 4.3% and 86.2% (not gated on CD45), respectively. The starting frequency of B cells in the non-lysed whole blood start sample above is 0.004% (data not shown). The purity of isolated cells is typically higher when processing samples that have an elevated starting frequency of B cells (e.g. CLL samples).

STEMCELL TECHNOLOGIES INC.’S QUALITY MANAGEMENT SYSTEM IS CERTIFIED TO ISO 13485. PRODUCTS ARE FOR RESEARCH USE ONLY AND NOT INTENDED FOR HUMAN OR ANIMAL DIAGNOSTIC OR THERAPEUTIC USES UNLESS OTHERWISE STATED.