EasySep™ Direct Human T Cell Isolation Kit
Immunomagnetic negative selection from whole blood kit
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EasySep™ Direct Human T Cell Isolation Kit -
Label for EasySep™ Direct Human T Cell Isolation 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
This cell separation kit can be used to isolate highly purified T cells directly from human whole blood by immunomagnetic negative selection.
This kit targets non-T 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 can directly be used for functional assays, flow cytometry or DNA/RNA extraction.
This kit targets non-T 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 can directly be used for functional assays, flow cytometry or DNA/RNA extraction.
Advantages:
• > 99.9% RBC depletion without the need for density gradient centrifugation, sedimentation, or lysis
• Up to 97% purity of isolated cells
• Fast, easy-to-use and column-free
• Isolated cells are untouched
• Up to 97% purity of isolated cells
• Fast, easy-to-use and column-free
• Isolated cells are untouched
Components:
- EasySep™ Direct Human T Cell Isolation Kit (Catalog #19661)
- EasySep™ Direct Human T Cell Isolation Cocktail, 2 x 2.5 mL
- EasySep™ Direct RapidSpheres™, 4 x 2.5 mL
- EasySep™ Direct Human T 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)
• RoboSep™-S (Catalog #21000)
Subtype:
Cell Isolation Kits
Cell Type:
T Cells
Species:
Human
Sample Source:
Whole Blood
Selection Method:
Negative
Application:
Cell Isolation
Brand:
EasySep; RoboSep
Area of Interest:
Cell Therapy; Immunology
Related Products
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EasySep™ Human T Cell Isolation Kit
8-Minute cell isolation kit using immunomagnetic negative selection -
Primary human cells, fresh
Labeling Antibodies
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Anti-Human CD4 Antibody, Clone OKT4
Mouse monoclonal IgG2b antibody against human, rhesus, cynomolgus CD4 -
Anti-Human CD8a Antibody, Clone RPA-T8
Mouse monoclonal IgG1 antibody against human, rhesus, cynomolgus CD8a -
Anti-Human CD3 Antibody, Clone UCHT1
Mouse (BALB/c) monoclonal IgG1 antibody against human, chimpanzee CD3
Scientific Resources
Product Documentation
Document Type
Product Name
Catalog #
Lot #
Language
19661RF
18F92420 or higher
Educational Materials
(15)
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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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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
Figure 1. Typical EasySep™ Direct Human T Cell Isolation Profile
Starting with human whole blood from normal healthy donors, the typical T cell (CD3+) content of the non-lysed final isolated fraction is 95.3 ± 1.4% (gated on CD45) or 94.9 ± 1.5% (not gated on CD45). In the example above, the T cell (CD3+) content of the lysed whole blood start sample and non-lysed final isolated fraction is 33.0% and 95.0% (gated on CD45), respectively, or 33.0% and 94.9% (not gated on CD45), respectively. The starting frequency of T cells in the non-lysed whole blood start sample above is 0.059% (data not shown).
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.