EasySep™ Direct Human CTC Enrichment Kit
Immunomagnetic negative selection from whole blood kit
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EasySep™ Direct Human CTC Enrichment Kit -
Label for EasySep™ Direct Human CTC Enrichment Kit
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Required Products
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Magnet for column-free immunomagnetic separation -
Dulbecco’s phosphate-buffered saline without calcium and magnesium -
Compatible antibodies for purity assessment of isolated cells
Overview
The EasySep™ Direct Human CTC Enrichment Kit can be used to enrich Circulating Tumor Cells (CTCs) directly from human whole blood by immunomagnetic negative selection without the need for either density gradient centrifugation or RBC lysis. Isolated cells are immediately ready for downstream analysis or assays.
Advantages:
• > 99.9% RBC depletion without the need for density gradient centrifugation, sedimentation, or lysis
• Up to 3 log depletion of normal hematopoietic cells
• Fast, easy-to-use and column-free
• Isolated cells are untouched
• Up to 3 log depletion of normal hematopoietic cells
• Fast, easy-to-use and column-free
• Isolated cells are untouched
Components:
- EasySep™ Direct Human CTC Enrichment Kit (Catalog #19657)
- EasySep™ Direct Human CTC Enrichment 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:
Cancer Cells and Cell Lines
Species:
Human
Sample Source:
Whole Blood
Selection Method:
Negative
Application:
Cell Isolation
Brand:
EasySep
Area of Interest:
Cancer Research; Immunology
Related Products
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RosetteSep™ CTC Enrichment Cocktail Containing Anti-CD36
Immunodensity negative selection cocktail -
RosetteSep™ CTC Enrichment Cocktail Containing Anti-CD56
Immunodensity negative selection cocktail
Labeling Antibodies
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Anti-Human Epithelial Cell Antibody, Clone 5E11.3.1
Mouse monoclonal IgG1 antibody against human epithelial cells -
Anti-Human CD326 (EpCAM) Antibody, Clone VU-1D9
Mouse monoclonal IgG1 antibody against human CD326 (EpCAM)
Scientific Resources
Product Documentation
Document Type
Product Name
Catalog #
Lot #
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Educational Materials
(11)
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.
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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Data and Publications
Data
Figure 1. Typical EasySep™ Direct Human CTC Enrichment Profile
Starting with human whole blood from healthy donors, spiked with approximately 1% of CAMA cells (epithelial tumor cell line), the typical CTC (epithelial cell+) content of non-lysed final enriched fraction is 79 ± 16 % (using the silver “Big Easy” EasySep™ Magnet; gated on DRAQ5™ for nucleated cells). Typically the log depletion of targeted CD45+ cells is 2.8 to 3.2. In the above example, CAMA cells were seeded into whole blood at a starting frequency of 0.98%. The CAMA cell (epithelial cell+) content of the enriched fraction is 92.02% with a 3.8 log depletion of CD45+ cells.
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.