New format, same high quality! You may notice that your kit contents and packaging look slightly different from previous orders. We are currently updating the format of select EasySep? Mouse kits to include a Mouse FcR blocker instead of Normal Rat Serum. With this change, all components will now be shipped in a single package, while providing the same cell isolation performance as before.
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Easily and efficiently isolate highly purified mouse T cells from single-cell suspensions of splenocytes or other tissues by immunomagnetic negative selection, with the EasySep? Mouse T Cell Isolation Kit. Widely used in published research for more than 20 years, EasySep? combines the specificity of monoclonal antibodies with the simplicity of a column-free magnetic system.
In this EasySep? negative selection procedure, unwanted cells are labeled with antibody complexes and magnetic particles. Unwanted cells expressing the following markers are targeted for removal: CD11b, CD45R, Ter119, CD49b, CD19, and CD24. The magnetically labeled cells are then separated from the untouched desired mouse T cells by using an EasySep? magnet and simply pouring or pipetting the desired cells into a new tube. Following magnetic cell isolation in as little as 15 minutes, the desired T cells are ready for downstream applications such as flow cytometry, culture, and cell-based experiments.
Learn more about how immunomagnetic EasySep? technology works or how to fully automate immunomagnetic cell isolation with RoboSep?. Explore additional products optimized for your workflow, including culture media, supplements, antibodies, and more.
Figure 1.Typical EasySep? Mouse T Cell Isolation Profile
Starting with mouse splenocytes, the T cell content (CD3+CD19-) of the isolated fraction is 96.6 ± 2.0% (mean ± SD), using the purple EasySep? magnet.
Figure 2.Cell Isolation Protocol Lengths
Typical time taken (in minutes) to isolate cells using select EasySep? kits.
Figure 3.ImmunoCult? Mouse T Cell Activator Kit Supports High Viability of Activated T Cells
Mouse T cells were isolated using EasySep? Mouse T Cell Isolation Kit (Catalog #19851), stimulated with ImmunoCult? Mouse T Cell Activator Kit (Catalog #100-1572), and cultured in IMDM + FBS formulation. Following 3 days of culture, the mean ± SD frequency of CD25+ cells was 91.9 ± 5.1% (n = 11) or 99.9 ± 0.1% (n = 5), when stimulated with ImmunoCult? Mouse CD3/CD28 T Cell Activator or ImmunoCult? Mouse CD3/CD28/CD2 T Cell Activator, respectively. Stimulated mouse T cells maintained expression levels of CD25 throughout the 7-day culture period.
Figure 4.Robust Expansion of EasySep?-Isolated Mouse T Cells Can Be Achieved Following Stimulation with ImmunoCult? Mouse T Cell Activator Kit
Mouse T cells isolated using EasySep? Mouse T Cell Isolation Kit (Catalog #19851) were expanded with ImmunoCult? Mouse T Cell Activator Kit (Catalog #100-1572) in IMDM + FBS formulation over 7 days. The number of viable cells was assessed every 2 - 3 days, and fresh medium supplemented with IL-2 was added. No additional ImmunoCult? Mouse T Cell Activator was added during the 7-day culture period. After 7 days in culture with ImmunoCult? Mouse CD3/CD28 T Cell Activator or ImmunoCult? Mouse CD3/CD28/CD2 T Cell Activator, stimulation resulted in a fold expansion of 23 ± 3.4 or 29.3 ± 4.8 (mean ± SEM, n = 6), respectively.
Figure 5.High Cell Proliferation is Observed in EasySep?-Isolated T cells After Stimulation with ImmunoCult? Mouse T Cell Activator
Mouse T cells isolated using EasySep? Mouse T Cell Isolation Kit (Catalog #19851) were labeled with CFDA-SE (Catalog #75003), stimulated with ImmunoCult? Mouse T Cell Activator Kit (Catalog #100-1572), and cultured in cultured in IMDM + FBS formulation. On Day 3, cells were harvested, stained with anti-mouse CD4 and CD8a antibodies, then measured by flow cytometry. Shown are CFDA-SE-labeled mouse T cells, gated on viable CD4+ (A) or CD8a+ (B) cells, cultured with no activator (top panel), with ImmunoCult? Mouse CD3/CD28 T Cell Activator (middle panel), or with ImmunoCult? Mouse CD3/CD28/CD2 T Cell Activator (bottom panel). Due to cell proliferation, the intensity of CFDA-SE signal is reduced by 50% for each cell division.
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.
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.
Bi-specific T cell-engaging antibody triggers protective immune memory and glioma microenvironment remodeling in immune-competent preclinical models
M. Zannikou et al.
Journal for Immunotherapy of Cancer 2025 Oct
Abstract
Bispecific T cell-engagers (BTEs) are engineered antibodies that redirect T cells to target antigen-expressing tumors. BTEs targeting tumor-specific antigens such as interleukin 13 receptor alpha 2 (IL13RA2) and epidermal growth factor receptor variant III (EGFRvIII) have been developed for glioblastoma (GBM). However, there is limited mechanistic understanding of the action of BTE since prior studies were mostly conducted in immunocompromised animal models. To close this gap, the function of BTEs was assessed in the immunosuppressive tumor microenvironment (TME) of orthotopic and genetically engineered mouse models (GEMM) with intact immune systems. Method: sA BTE that bridges CD3 epsilon on murine T cells to IL13RA2-positive GBM cells was developed, and the therapeutic mechanism was investigated in immunocompetent mouse models of GBM. Multicolor flow cytometry, single-cell RNA sequencing (scRNA-seq), multiplex immunofluorescence, and multiparametric MRI across multiple preclinical models of GBM were used to evaluate the mechanism of action and response. Results: BTE-mediated interactions between murine T cells and GBM cells triggered T cell activation and antigen-dependent killing of GBM cells. BTE treatment significantly extended the survival of mice bearing IL13RA2-expressing orthotopic glioma and de novo forming GBM in the GEMM. Quantified parametric MRI validated the survival data, showing a reduction in glioma volume and decreased glioma viability. Flow cytometric and scRNA-seq analyses of the TME revealed robust increases in activated and memory T cells and decreases in immunosuppressive myeloid cells within the brains of mice following BTE treatment. Conclusions: Our data demonstrate that the survival benefits of BTEs in preclinical models of glioma are due to the ability to engage the host immune system in direct killing, induction of immunological memory, and modulation of the TME. These findings provide a deeper insight into the mechanism of BTE actions in GBM.
A CARMIL2 gain-of-function mutation suffices to trigger most CD28 costimulatory functions in vivo
The Journal of Experimental Medicine 2025 May
Abstract
Zhang et al. demonstrate that the expression of a mutated CARMIL2 protein in CD28-deficient mice induces most of the developmental and functional consequences known to result from CD28 costimulation and in turn triggers potent tumor-specific T cell responses resistant to PD-1 and CTLA-4 blockade. Naive T cell activation requires both TCR and CD28 signals. The CARMIL2 cytosolic protein enables CD28-dependent activation of the NF-κB transcription factor via its ability to link CD28 to the CARD11 adaptor protein. Here, we developed mice expressing a mutation named Carmil2QE and mimicking a mutation found in human T cell malignancies. Naive T cells from Carmil2QE mice contained preformed CARMIL2QE-CARD11 complexes in numbers comparable to those assembling in wild-type T cells after CD28 engagement. Such ready-made CARMIL2QE-CARD11 complexes also formed in CD28-deficient mice where they unexpectedly induced most of the functions that normally result from CD28 engagement in a manner that remains antigen-dependent. In turn, tumor-specific T cells expressing Carmil2QE do not require CD28 engagement and thereby escape to both PD-1 and CTLA-4 inhibition. In conclusion, we uncovered the overarching role played by CARMIL2-CARD11 signals among those triggered by CD28 and exploited them to induce potent solid tumor–specific T cell responses in the absence of CD28 ligands and immune checkpoint inhibitors.
Adoptive Transfer of T Cells as a Potential Therapeutic Approach in the Bleomycin‐Injured Mouse Lung
The Journal of Gene Medicine 2025 Mar
Abstract
ABSTRACTBackgroundIdiopathic pulmonary fibrosis (IPF) is a lethal disease with an unknown etiology and complex pathophysiology that are not fully understood. The disease involves intricate cellular interplay, particularly among various immune cells. Currently, there is no treatment capable of reversing the fibrotic process or aiding lung regeneration. Hepatocyte growth factor (HGF) has demonstrated antifibrotic properties, whereas the adoptive transfer of modified T cells is a well‐established treatment for various malignancies. We aimed to understand the dynamics of T cells in the progression of lung fibrosis and to study the therapeutic benefit of adoptive T cell transfer in a bleomycin‐injured mouse lung (BLM) model.MethodsT cells were isolated from the spleen of na?ve mice and transfected in?vitro with mouse HGF plasmid and were administered intratracheally to the mice lungs 7?days post‐bleomycin injury to the lung. Lung tissue and bronchoalveolar lavage were collected and analyzed using flow cytometry, histology, qRT‐PCR, ELISA, and hydroxyproline assay.ResultsOur findings demonstrate the successful T cell therapy of bleomycin‐induced lung injury through the adoptive transfer of HGF‐transfected T cells in mice. This treatment resulted in decreased collagen deposition and a balancing of immune cell exhaustion and cytokine homeostasis compared with untreated controls. In?vitro testing showed enhanced apoptosis in myofibroblasts induced by HGF‐overexpressing T cells.ConclusionsTaken together, our data highlight the great potential of adoptive T cell transfer as an emerging therapy to counteract lung fibrosis. This study explores the potential of T cells as a therapeutic strategy against idiopathic pulmonary fibrosis (IPF), a progressive lung disease for which there is currently no treatment to reverse fibrosis or restore normal lung function. To investigate an innovative approach using adoptive T cell transfer, T cells isolated from healthy mice were genetically modified to carry a plasmid containing hepatocyte growth factor (HGF). The modified cells were delivered directly into the airways of mice with bleomycin‐induced lung fibrosis. The results showed a significant reduction in fibrotic scarring, improved immune regulation, and increased apoptosis of pathogenic myofibroblasts. These results highlight the potential of HGF‐engineered T cells as a promising therapeutic approach to combat IPF.
Hamster (Armenian) monoclonal IgG1 antibody against mouse CD3e
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EasySep? Mouse T Cell Isolation Kit
New format, same high quality! You may notice that your kit contents and packaging look slightly different from previous orders. We are currently updating the format of select EasySep? Mouse kits to include a Mouse FcR blocker instead of Normal Rat Serum. With this change, all components will now be shipped in a single package, while providing the same cell isolation performance as before.
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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 海角破解版, REFER TO WWW.海角破解版.COM/COMPLIANCE.