EasySep™ Mouse Memory CD4+ T Cell Isolation Kit
27.5-Minute cell isolation kit using immunomagnetic negative selection
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Required Products
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EasyEights™ EasySep™ MagnetMagnet for column-free immunomagnetic separation
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EasySep™ BufferCell separation buffer
Overview
The EasySep™ Mouse Memory CD4+ T Cell Isolation Kit is designed to isolate pan-memory CD4+ T cells from single-cell suspensions of splenocytes or other tissues by negative selection. Unwanted cells are targeted for removal with biotinylated antibodies directed against non-memory CD4+ T cells (CD8, CD11b, CD11c, CD19, CD24, CD45R, CD45RB, CD49b, CD105 and TCRγ/δ) and streptavidin-coated magnetic particles (RapidSpheres™ ). Labeled cells are separated using an EasySep™ magnet without the use of columns. Desired cells are poured off into a new tube.
Learn more about our next-generation EasySep™ mouse cell isolation kits, featuring RapidSphere™ technology.
Learn more about our next-generation EasySep™ mouse cell isolation kits, featuring RapidSphere™ technology.
Advantages
• Fast, easy-to-use and column-free
• Up to 96% purity
• Untouched, viable cells
• Up to 96% purity
• Untouched, viable cells
Components
- EasySep™ Mouse Memory CD4+ T Cell Isolation Kit (Catalog #19767)
- EasySep™ Mouse Memory CD4+ T Cell Isolation Cocktail, 0.5 mL
- EasySep™ Streptavidin RapidSpheres™ 50001, 2 x 1 mL
- Normal Rat Serum, 2 mL
- RoboSep™ Mouse Memory CD4+ T Cell Isolation Kit (Catalog #19767RF)
- EasySep™ Mouse Memory CD4+ T Cell Isolation Cocktail, 0.5 mL
- EasySep™ Streptavidin RapidSpheres™ 50001, 2 x 1 mL
- Normal Rat Serum, 2 mL
- RoboSep™ Buffer (Catalog #20104)
- RoboSep™ Filter Tips (Catalog #20125)
Magnet Compatibility
• EasySep™ Magnet (Catalog #18000)
• “The Big Easy” EasySep™ Magnet (Catalog #18001)
• EasyEights™ EasySep™ Magnet (Catalog #18103)
• RoboSep™-S (Catalog #21000)
Subtype
Cell Isolation Kits
Cell Type
T Cells, T Cells, CD4+
Species
Mouse
Sample Source
Other, Spleen
Selection Method
Negative
Application
Cell Isolation
Brand
EasySep, RoboSep
Area of Interest
Immunology
Scientific Resources
Product Documentation
| Document Type | Product Name | Catalog # | Lot # | Language |
|---|---|---|---|---|
|
Document Type
Product Information Sheet
|
Product Name
EasySep™ Mouse Memory CD4+ T Cell Isolation Kit
|
Catalog #
19767 |
Lot #
All |
Language
English |
|
Document Type
Product Information Sheet
|
Product Name
RoboSep™ Mouse Memory CD4+ T Cell Isolation Kit
|
Catalog #
19767RF |
Lot #
All |
Language
English |
|
Document Type
Safety Data Sheet 1
|
Product Name
EasySep™ Mouse Memory CD4+ T Cell Isolation Kit
|
Catalog #
19767 |
Lot #
All |
Language
English |
|
Document Type
Safety Data Sheet 2
|
Product Name
EasySep™ Mouse Memory CD4+ T Cell Isolation Kit
|
Catalog #
19767 |
Lot #
All |
Language
English |
|
Document Type
Safety Data Sheet 3
|
Product Name
EasySep™ Mouse Memory CD4+ T Cell Isolation Kit
|
Catalog #
19767 |
Lot #
All |
Language
English |
|
Document Type
Safety Data Sheet 1
|
Product Name
RoboSep™ Mouse Memory CD4+ T Cell Isolation Kit
|
Catalog #
19767RF |
Lot #
All |
Language
English |
|
Document Type
Safety Data Sheet 2
|
Product Name
RoboSep™ Mouse Memory CD4+ T Cell Isolation Kit
|
Catalog #
19767RF |
Lot #
All |
Language
English |
|
Document Type
Safety Data Sheet 3
|
Product Name
RoboSep™ Mouse Memory CD4+ T Cell Isolation Kit
|
Catalog #
19767RF |
Lot #
All |
Language
English |
Educational Materials (9)
Brochure
Mouse Cell Isolation
Brochure
Tools For Your Immunology Research
Brochure
EasySep™ Cell Separation Technology
Wallchart
Antigen Processing and Presentation
Wallchart
Human Immune Cytokines
Video
1:13
Isolate Cells with a Simple Pour-Off: EasySep™ Cell Separation Technology
Video
0:57
Simultaneous Cell Isolation from Multiple Samples Using the EasyEights™ EasySep™ Magnet
Video
0:01
EasySep™ Mouse Cell Isolation Kits: How Much Time Will You Save?
Video
1:57
How EasySep™ Magnetic Cell Separation Technology Works: Fast and Easy Cell Isolation
Frequently Asked Question
Can EasySep™ Streptavidin RapidSpheres™ be used for either positive or negative selection?
Currently, EasySep™ Streptavidin RapidSphere™ kits are only available for negative selection and work by targeting and removing unwanted cells.
How does the separation work?
Streptavidin RapidSphere™ magnetic particles are crosslinked to unwanted cells using biotinylated antibodies. When placed in the EasySep™ Magnet, labeled cells migrate to the wall of the tube. The unlabeled cells are then poured off into a new tube.
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™ Streptavidin RapidSphere™ separations be automated?
Yes. RoboSep™, the fully automated cell separator, automates all EasySep™ labeling and cell separation steps.
Are cells isolated using EasySep™ RapidSphere™ products FACS-compatible?
Yes. Desired cells are unlabeled and ready to use in downstream applications, such as FACS analysis.
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.
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.
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.
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.
Which cell separation kits are compatible with the EasyPlate™ EasySep™ magnet?
The EasyPlate™-compatible human EasySep™ kits are:
19051 (T Cells), 19052 (CD4 T cells), 19157 (Memory CD4 T Cells), 19053 (CD8 T Cells), 19054 (B Cells), 19254 (Naïve B cells), 19055 (NK Cells), 19058 (Monocytes without CD16 depletion), 19059 (Monocytes)
The EasyPlate™-compatible mouse EasySep™ kits are:
19751 (T Cells), 19752 (CD4 T cells), 19753 (CD8 T Cells), 19754 (B Cells), 19755 (NK Cells - please contact Tech Support)
19051 (T Cells), 19052 (CD4 T cells), 19157 (Memory CD4 T Cells), 19053 (CD8 T Cells), 19054 (B Cells), 19254 (Naïve B cells), 19055 (NK Cells), 19058 (Monocytes without CD16 depletion), 19059 (Monocytes)
The EasyPlate™-compatible mouse EasySep™ kits are:
19751 (T Cells), 19752 (CD4 T cells), 19753 (CD8 T Cells), 19754 (B Cells), 19755 (NK Cells - please contact Tech Support)
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.
Research Area
Workflow Stages
Data and Publications
Data
Figure 1. Typical EasySet™ Mouse Memory CD4+ T Cell Isolation Profile
Starting with mouse splenocytes, the memory CD4+ T cell (CD4+CD62L high CD44 high/int central memory and CD4+CD62L-CD44 high/int effector memory) content of the isolated fraction typically ranges from 78 - 96%. In the example above, the final purities of the start and isolated fractions are 5.3% and 90.2%, respectively.
Publications (1)
Nature communications 2020
T cell-intrinsic role for Nod2 in protection against Th17-mediated uveitis.
Abstract
Abstract
Mutations in nucleotide-binding oligomerization domain-containing protein 2 (NOD2) cause Blau syndrome, an inflammatory disorder characterized by uveitis. The antimicrobial functions of Nod2 are well-established, yet the cellular mechanisms by which dysregulated Nod2 causes uveitis remain unknown. Here, we report a non-conventional, T cell-intrinsic function for Nod2 in suppression of Th17 immunity and experimental uveitis. Reconstitution of lymphopenic hosts with Nod2-/- CD4+ T cells or retina-specific autoreactive CD4+ T cells lacking Nod2 reveals a T cell-autonomous, Rip2-independent mechanism for Nod2 in uveitis. In naive animals, Nod2 operates downstream of TCR ligation to suppress activation of memory CD4+ T cells that associate with an autoreactive-like profile involving IL-17 and Ccr7. Interestingly, CD4+ T cells from two Blau syndrome patients show elevated IL-17 and increased CCR7. Our data define Nod2 as a T cell-intrinsic rheostat of Th17 immunity, and open new avenues for T cell-based therapies for Nod2-associated disorders such as Blau syndrome.
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 STEMCELL, REFER TO WWW.STEMCELL.COM/COMPLIANCE.
