EasySep™ Mouse T Cell Isolation Kit

15-Minute cell isolation kit using immunomagnetic negative selection

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.

EasySep™ Mouse T Cell Isolation Kit

15-Minute cell isolation kit using immunomagnetic negative selection

From: 776 USD
Catalog #
(Select a product)
15-Minute cell isolation kit using immunomagnetic negative selection
Add to Wish List

Product Advantages


  • Fast and easy-to-use

  • Up to 99% purity

  • No columns required

  • Untouched, viable cells

What's Included

  • EasySep™ Mouse T Cell Isolation Kit (Catalog #19851)
    • EasySep™ Mouse T Cell Isolation Cocktail, 0.5 mL
    • EasySep™ Streptavidin RapidSpheres™ 50001, 1 mL
    • EasySep™ FcR Mouse Blocker, 0.2 mL
  • RoboSep™ Mouse T Cell Isolation Kit (Catalog #19851RF)
    • EasySep™ Mouse T Cell Isolation Cocktail, 0.5 mL
    • EasySep™ Streptavidin RapidSpheres™ 50001, 1.0 mL
    • EasySep™ FcR Mouse Blocker, 0.2 mL
    • RoboSep™ Buffer (Catalog #20104)
    • RoboSep™ Filter Tips (Catalog #20125)
Products for Your Protocol

Overview

The EasySep™ Mouse 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-T cells 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.

This product replaces the EasySep™ Mouse T Cell Enrichment Kit (Catalog #19751) for even faster cell isolations.
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
Species
Mouse
Sample Source
Other, Spleen
Selection Method
Negative
Application
Cell Isolation
Brand
EasySep, RoboSep
Area of Interest
Immunology

Data Figures

Typical EasySep™ Mouse T Cell Isolation Profile

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.

Cell Isolation Protocol Lengths

Figure 2.Cell Isolation Protocol Lengths

Typical time taken (in minutes) to isolate cells using select EasySep™ kits.

ImmunoCult™ Mouse T Cell Activator Kit Supports High Viability of Activated T Cells

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.

Robust Expansion of EasySep™-Isolated Mouse T Cells Can Be Achieved Following Stimulation with ImmunoCult™ Mouse T Cell Activator Kit

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.

High Cell Proliferation is Observed in EasySep™-Isolated T cells After Stimulation with ImmunoCult™ Mouse T Cell Activator

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.

Protocols and Documentation

Find supporting information and directions for use in the Product Information Sheet or explore additional protocols below.

Document Type
Product Name
Catalog #
Lot #
Language
Catalog #
19851
Lot #
1000120297 or lower
Language
English
Catalog #
19851
Lot #
1000120298 or higher
Language
English
Catalog #
19851RF
Lot #
1000120298 or higher
Language
English
Catalog #
19851RF
Lot #
1000120297 or lower
Language
English
Document Type
Safety Data Sheet 1
Catalog #
19851
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Catalog #
19851
Lot #
All
Language
English
Document Type
Safety Data Sheet 3
Catalog #
19851
Lot #
All
Language
English
Document Type
Safety Data Sheet 4
Catalog #
19851
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Catalog #
19851RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Catalog #
19851RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 3
Catalog #
19851RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 4
Catalog #
19851RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 5
Catalog #
19851RF
Lot #
All
Language
English

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.

Resources and Publications

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.

Publications (18)

Bioluminescence for in vivo detection of cell-type-specific inflammation in a mouse model of uveitis. S. John et al. Scientific reports 2020 jul

Abstract

This study reports the use of cell-type-specific in vivo bioluminescence to measure intraocular immune cell population dynamics during the course of inflammation in a mouse model of uveitis. Transgenic lines expressing luciferase in inflammatory cell subsets (myeloid cells, T cells, and B cells) were generated and ocular bioluminescence was measured serially for 35 days following uveitis induction. Ocular leukocyte populations were identified using flow cytometry and compared to the ocular bioluminescence profile. Acute inflammation is neutrophilic (75{\%} of ocular CD45 + cells) which is reflected by a significant increase in ocular bioluminescence in one myeloid reporter line on day 2. By day 7, the ocular T cell population increases to 50{\%} of CD45 + cells, leading to a significant increase in ocular bioluminescence in the T cell reporter line. While initially negligible ({\textless} 1{\%} of CD45 + cells), the ocular B cell population increases to {\textgreater} 4{\%} by day 35. This change is reflected by a significant increase in the ocular bioluminescence of the B cell reporter line starting on day 28. Our data demonstrates that cell-type-specific in vivo bioluminescence accurately detects changes in multiple intraocular immune cell populations over time in experimental uveitis. This assay could also be useful in other inflammatory disease models.
VEGF-C-driven lymphatic drainage enables immunosurveillance of brain tumours. E. Song et al. Nature 2020

Abstract

Immune surveillance against pathogens and tumours in the central nervous system is thought to be limited owing to the lack of lymphatic drainage. However, the characterization of the meningeal lymphatic network has shed light on previously unappreciated ways that an immune response can be elicited to antigens that are expressed in the brain1-3. Despite progress in our understanding of the development and structure of the meningeal lymphatic system, the contribution of this network in evoking a protective antigen-specific immune response in the brain remains unclear. Here, using a mouse model of glioblastoma, we show that the meningeal lymphatic vasculature can be manipulated to mount better immune responses against brain tumours. The immunity that is mediated by CD8 T cells to the glioblastoma antigen is very limited when the tumour is confined to the central nervous system, resulting in uncontrolled tumour growth. However, ectopic expression of vascular endothelial growth factor C (VEGF-C) promotes enhanced priming of CD8 T cells in the draining deep cervical lymph nodes, migration of CD8 T cells into the tumour, rapid clearance of the glioblastoma and a long-lasting antitumour memory response. Furthermore, transfection of an mRNA construct that expresses VEGF-C works synergistically with checkpoint blockade therapy to eradicate existing glioblastoma. These results reveal the capacity of VEGF-C to promote immune surveillance of tumours, and suggest a new therapeutic approach to treat brain tumours.
ICOS Is an Indicator of T-cell-Mediated Response to Cancer Immunotherapy. Z. Xiao et al. Cancer research 2020

Abstract

Immunotherapy is innovating clinical cancer management. Nevertheless, only a small fraction of patient's benefit from current immunotherapies. To improve clinical management of cancer immunotherapy, it is critical to develop strategies for response monitoring and prediction. In this study, we describe inducible T-cell costimulator (ICOS) as a conserved mediator of immune response across multiple therapy strategies. ICOS expression was evaluated by flow cytometry, 89Zr-DFO-ICOS mAb PET/CT imaging was performed on Lewis lung cancer models treated with different immunotherapy strategies, and the change in tumor volume was used as a read-out for therapeutic response. ImmunoPET imaging of ICOS enabled sensitive and specific detection of activated T cells and early benchmarking of immune response. A STING (stimulator of interferon genes) agonist was identified as a promising therapeutic approach in this manner. The STING agonist generated significantly stronger immune responses as measured by ICOS ImmunoPET and delayed tumor growth compared with programmed death-1 checkpoint blockade. More importantly, ICOS ImmunoPET enabled early and robust prediction of therapeutic response across multiple treatment regimens. These data show that ICOS is an indicator of T-cell-mediated immune response and suggests ICOS ImmunoPET as a promising strategy for monitoring, comparing, and predicting immunotherapy success in cancer. SIGNIFICANCE: ICOS ImmunoPET is a promising strategy to noninvasively predict and monitor immunotherapy response.See related commentary by Choyke, p. 2975.
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.