EasySep™ Mouse Pan-Naïve T Cell Isolation Kit
15-Minute cell isolation kit using immunomagnetic negative selection
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EasySep™ Mouse Pan-Naïve T Cell Isolation Kit -
RoboSep™ Mouse Pan-Naïve T Cell Isolation Kit -
Label for EasySep™ Mouse Pan-Naïve T Cell Isolation Kit
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Required Products
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Magnet for column-free immunomagnetic separation -
Cell separation buffer -
Compatible antibodies for purity assessment of isolated cells
Overview
The EasySep™ Mouse Pan-Naïve T Cell Isolation Kit is designed to isolate 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-naïve T cells (CD11b, CD19, CD24, CD25, CD44, CD45R/B220, CD49b, TER119) and streptavidin-coated magnetic particles. Labeled cells are separated using an EasySep™ magnet without the use of columns. Desired cells are poured off into a new tube.
Click here to learn about our next-generation EasySep™ mouse cell isolation kits, featuring RapidSphere™ technology.
Click here to learn about our next-generation EasySep™ mouse cell isolation kits, featuring RapidSphere™ technology.
Advantages:
• Fast and easy-to-use
• Up to 97% purity
• No columns required
• Untouched, viable cells
• Up to 97% purity
• No columns required
• Untouched, viable cells
Components:
- EasySep™ Mouse Pan-Naïve T Cell Isolation Kit (Catalog #19848)
- EasySep™ Mouse T Cell Isolation Cocktail, 0.5 mL
- EasySep™ Mouse Memory T Cell Depletion Cocktail, 0.5 mL
- EasySep™ Streptavidin RapidSpheres™ 50001, 1 mL
- Normal Rat Serum, 2 mL
- RoboSep™ Mouse Pan-Naïve T Cell Isolation Kit (Catalog #19848RF)
- EasySep™ Mouse T Cell Isolation Cocktail, 0.5 mL
- EasySep™ Mouse Memory T Cell Depletion Cocktail, 0.5 mL
- EasySep™ Streptavidin RapidSpheres™ 50001, 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)
• EasyPlate™ EasySep™ Magnet (Catalog 18102)
• EasyEights™ EasySep™ Magnet (Catalog #18103)
• RoboSep™-S (Catalog #21000)
Subtype:
Cell Isolation Kits
Cell Type:
T Cells
Species:
Mouse
Sample Source:
Other; Spleen
Selection Method:
Negative
Application:
Cell Isolation
Brand:
EasySep; RoboSep
Area of Interest:
Immunology
Related Products
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EasySep™ Mouse Naïve CD4+ T Cell Isolation Kit
15-Minute cell isolation kit using immunomagnetic negative selection -
EasySep™ Mouse CD4+ T Cell Isolation Kit
15-Minute cell isolation kit using immunomagnetic negative selection -
EasySep™ Mouse T Cell Isolation Kit
15-Minute cell isolation kit using immunomagnetic negative selection
Labeling Antibodies
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Anti-Mouse CD62L (L-Selectin) Antibody, Clone MEL-14
Rat monoclonal IgG2a antibody against mouse CD62L (L-selectin) -
Anti-Mouse CD3e Antibody, Clone 145-2C11
Hamster (Armenian) monoclonal IgG1 antibody against mouse CD3e
Scientific Resources
Product Documentation
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Catalog #
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Educational Materials
(9)
Frequently Asked Questions
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.
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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
Figure 1. Typical EasySep™ Mouse Pan-Naïve T Cell Isolation Profile
Starting with mouse splenocytes from an uninfected mouse, the pan-naïve T cell (CD3+CD44-/lowCD62Lhigh) content of the isolated fraction typically ranges from 90 - 97%. In the example above, the final purities of the start and isolated fractions are 18.5% and 94.5%, respectively.
Publications
(2)
Nature 2019 apr
Nuclear positioning facilitates amoeboid migration along the path of least resistance.
Abstract
Abstract
During metazoan development, immune surveillance and cancer dissemination, cells migrate in complex three-dimensional microenvironments1-3. These spaces are crowded by cells and extracellular matrix, generating mazes with differently sized gaps that are typically smaller than the diameter of the migrating cell4,5. Most mesenchymal and epithelial cells and some-but not all-cancer cells actively generate their migratory path using pericellular tissue proteolysis6. By contrast, amoeboid cells such as leukocytes use non-destructive strategies of locomotion7, raising the question how these extremely fast cells navigate through dense tissues. Here we reveal that leukocytes sample their immediate vicinity for large pore sizes, and are thereby able to choose the path of least resistance. This allows them to circumnavigate local obstacles while effectively following global directional cues such as chemotactic gradients. Pore-size discrimination is facilitated by frontward positioning of the nucleus, which enables the cells to use their bulkiest compartment as a mechanical gauge. Once the nucleus and the closely associated microtubule organizing centre pass the largest pore, cytoplasmic protrusions still lingering in smaller pores are retracted. These retractions are coordinated by dynamic microtubules; when microtubules are disrupted, migrating cells lose coherence and frequently fragment into migratory cytoplasmic pieces. As nuclear positioning in front of the microtubule organizing centre is a typical feature of amoeboid migration, our findings link the fundamental organization of cellular polarity to the strategy of locomotion.
The Journal of Immunology 2015
Skin Metabolites Define a New Paradigm in the Localization of Skin Tropic Memory T Cells
Abstract
Abstract
The localization of memory T cells to human skin is essential for long-term immune surveillance and the maintenance of barrier integrity. The expression of CCR8 during naive T cell activation is controlled by skin-specific factors derived from epidermal keratinocytes and not by resident dendritic cells. In this study, we show that the CCR8-inducing factors are heat stable and protease resistant and include the vitamin D3 metabolite 1α,25-dihydroxyvitamin D3 and PGE2. The effect of either metabolite alone on CCR8 expression was weak, whereas their combination resulted in robust CCR8 expression. Elevation of intracellular cAMP was essential because PGE2 could be substituted with the adenylyl cyclase agonist forskolin, and CCR8 expression was sensitive to protein kinase A inhibition. For effective induction, exposure of naive T cells to these epidermal factors needed to occur either prior to or during T cell activation even though CCR8 was only detected 4-5 d later in proliferating T cells. The importance of tissue environments in maintaining cellular immune surveillance networks within distinct healthy tissues provides a paradigm shift in adaptive immunity. Epidermal-derived vitamin D3 metabolites and PGs provide an essential cue for the localization of CCR8(+) immune surveillance T cells within healthy human skin.
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.