EasySep™ Mouse CD8+ T Cell Isolation Kit
17.5-Minute cell isolation kit using immunomagnetic negative selection
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EasySep™ Mouse CD8+ T Cell Isolation Kit -
RoboSep™ Mouse CD8+ T Cell Isolation Kit -
Label for EasySep™ Mouse CD8+ 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 CD8+ T Cell Isolation Kit is designed to isolate CD8+ 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-CD8+ T cells 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.
This product replaces the EasySep™ Mouse CD8+ T Cell Enrichment Kit (Catalog #19753) for even faster cell isolations.
This product replaces the EasySep™ Mouse CD8+ T Cell Enrichment Kit (Catalog #19753) for even faster cell isolations.
Advantages:
• Fast and easy-to-use
• Up to 95% purity
• No columns required
• Untouched, viable cells
• Up to 95% purity
• No columns required
• Untouched, viable cells
Components:
- EasySep™ Mouse CD8+ T Cell Isolation Kit (Catalog #19853)
- EasySep™ Mouse CD8+ T Cell Isolation Cocktail, 0.5 mL
- EasySep™ Streptavidin RapidSpheres™ 50001, 2 x 1 mL
- Normal Rat Serum, 2 mL
- RoboSep™ Mouse CD8+ T Cell Isolation Kit (Catalog #19853RF)
- EasySep™ Mouse CD8+ 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)
• EasyPlate™ EasySep™ Magnet (Catalog 18102)
• EasyEights™ EasySep™ Magnet (Catalog #18103)
• RoboSep™-S (Catalog #21000)
Subtype:
Cell Isolation Kits
Cell Type:
T Cells; T Cells, CD8+
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 CD8a Positive Selection Kit II
Immunomagnetic positive selection kit -
EasySep™ Mouse T Cell Isolation Kit
15-Minute cell isolation kit using immunomagnetic negative selection -
EasySep™ Mouse Naïve CD8+ T Cell Isolation Kit
17.5-Minute cell isolation kit using immunomagnetic negative selection
Labeling Antibodies
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Anti-Mouse CD8a Antibody, Clone 53-6.7
Rat monoclonal IgG2a antibody against mouse, toad CD8a
Scientific Resources
Product Documentation
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Educational Materials
(15)
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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.
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Workflow Stages for
Workflow Stages
Data and Publications
Data
Figure 1. Typical EasySep™ Mouse CD8+ T Cell Isolation Profile
Starting with mouse splenocytes, the CD8+ T cell content of the isolated fraction typically ranges from 87 - 95%.
Publications
(26)
Nature 2019 may
CD8+ T cells regulate tumour ferroptosis during cancer immunotherapy.
Abstract
Abstract
Cancer immunotherapy restores or enhances the effector function of CD8+ T cells in the tumour microenvironment1,2. CD8+ T cells activated by cancer immunotherapy clear tumours mainly by inducing cell death through perforin-granzyme and Fas-Fas ligand pathways3,4. Ferroptosis is a form of cell death that differs from apoptosis and results from iron-dependent accumulation of lipid peroxide5,6. Although it has been investigated in vitro7,8, there is emerging evidence that ferroptosis might be implicated in a variety of pathological scenarios9,10. It is unclear whether, and how, ferroptosis is involved in T cell immunity and cancer immunotherapy. Here we show that immunotherapy-activated CD8+ T cells enhance ferroptosis-specific lipid peroxidation in tumour cells, and that increased ferroptosis contributes to the anti-tumour efficacy of immunotherapy. Mechanistically, interferon gamma (IFNgamma) released from CD8+ T cells downregulates the expression of SLC3A2 and SLC7A11, two subunits of the glutamate-cystine antiporter system xc-, impairs the uptake of cystine by tumour cells, and as a consequence, promotes tumour cell lipid peroxidation and ferroptosis. In mouse models, depletion of cystine or cysteine by cyst(e)inase (an engineered enzyme that degrades both cystine and cysteine) in combination with checkpoint blockade synergistically enhanced T cell-mediated anti-tumour immunity and induced ferroptosis in tumour cells. Expression of system xc- was negatively associated, in cancer patients, with CD8+ T cell signature, IFNgamma expression, and patient outcome. Analyses of human transcriptomes before and during nivolumab therapy revealed that clinical benefits correlate with reduced expression of SLC3A2 and increased IFNgamma and CD8. Thus, T cell-promoted tumour ferroptosis is an anti-tumour mechanism, and targeting this pathway in combination with checkpoint blockade is a potential therapeutic approach.
Science (New York, N.Y.) 2019
LMBR1L regulates lymphopoiesis through Wnt/beta-catenin signaling.
Abstract
Abstract
Precise control of Wnt signaling is necessary for immune system development. In this study, we detected severely impaired development of all lymphoid lineages in mice, resulting from an N-ethyl-N-nitrosourea-induced mutation in the limb region 1-like gene (Lmbr1l), which encodes a membrane-spanning protein with no previously described function in immunity. The interaction of LMBR1L with glycoprotein 78 (GP78) and ubiquitin-associated domain-containing protein 2 (UBAC2) attenuated Wnt signaling in lymphocytes by preventing the maturation of FZD6 and LRP6 through ubiquitination within the endoplasmic reticulum and by stabilizing destruction complex" proteins. LMBR1L-deficient T cells exhibited hallmarks of Wnt/beta-catenin activation and underwent apoptotic cell death in response to proliferative stimuli. LMBR1L has an essential function during lymphopoiesis and lymphoid activation acting as a negative regulator of the Wnt/beta-catenin pathway."
Mucosal immunology 2018 NOV
GITRL on inflammatory antigen presenting cells in the lung parenchyma provides signal 4 for T-cell accumulation and tissue-resident memory T-cell formation.
Abstract
Abstract
T-cell responses in the lung are critical for protection against respiratory pathogens. TNFR superfamily members play important roles in providing survival signals to T cells during respiratory infections. However, whether these signals take place mainly during priming in the secondary lymphoid organs and/or in the peripheral tissues remains unknown. Here we show that under conditions of competition, GITR provides a T-cell intrinsic advantage to both CD4 and CD8 effector T cells in the lung tissue, as well as for the formation of CD4 and CD8 tissue-resident memory T cells during respiratory influenza infection in mice. In contrast, under non-competitive conditions, GITR has a preferential effect on CD8 over CD4 T cells. The nucleoprotein-specific CD8 T-cell response partially compensated for GITR deficiency by expansion of higher affinity T cells; whereas, the polymerase-specific response was less flexible and more GITR dependent. Following influenza infection, GITR is expressed on lung T cells and GITRL is preferentially expressed on lung monocyte-derived inflammatory antigen presenting cells. Accordingly, we show that GITR+/+ T cells in the lung parenchyma express more phosphorylated-ribosomal protein S6 than their GITR-/- counterparts. Thus, GITR signaling within the lung tissue critically regulates effector and tissue-resident memory T-cell accumulation.
Cell 2018 NOV
Protein Barcodes Enable High-Dimensional Single-Cell CRISPR Screens.
Abstract
Abstract
CRISPR pools are being widely employed to identify gene functions. However, current technology, which utilizes DNA as barcodes, permits limited phenotyping and bulk-cell resolution. To enable novel screening capabilities, we developed a barcoding system operating at the protein level. We synthesized modules encoding triplet combinations of linear epitopes to generate {\textgreater}100 unique protein barcodes (Pro-Codes). Pro-Code-expressing vectors were introduced into cells and analyzed by CyTOF mass cytometry. Using just 14 antibodies, we detected 364 Pro-Code populations; establishing the largest set of protein-based reporters. By pairing each Pro-Code with a different CRISPR, we simultaneously analyzed multiple phenotypic markers, including phospho-signaling, on dozens of knockouts. Pro-Code/CRISPR screens found two interferon-stimulated genes, the immunoproteasome component Psmb8 and a chaperone Rtp4, are important for antigen-dependent immune editing of cancer cells and identified Socs1 as a negative regulator of Pd-l1. The Pro-Code technology enables simultaneous high-dimensional protein-level phenotyping of 100s of genes with single-cell resolution.
The Journal of clinical investigation 2018 NOV
Type I IFN blockade uncouples immunotherapy-induced antitumor immunity and autoimmune toxicity.
Abstract
Abstract
Despite showing success in treating melanoma and haematological malignancies, adoptive cell therapy (ACT) has generated only limited effects in solid tumors. This is, in part, due to a lack of specific antigen targets, poor trafficking/infiltration and immunosuppression in the tumor microenvironment. In this study, we combined ACT with oncolytic virus vaccines (OVV) to drive expansion and tumor infiltration of transferred antigen-specific T cells, and demonstrated that the combination is highly potent for the eradication of established solid tumors. Consistent with other successful immunotherapies, this approach elicited severe autoimmune consequence when the antigen targeted was a self-protein. However, modulation of IFN$\alpha$/$\beta$ signaling, either by functional blockade or rational choice of an OVV backbone, ameliorated autoimmune side effects without compromising antitumor efficacy. Our study uncovers a pathogenic role for IFN$\alpha$/$\beta$ in facilitating autoimmune toxicity during cancer immunotherapy and offers a safe and powerful combinatorial regimen with immediate translational applications.
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.