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EasySep™ Mouse Naïve CD4+ T Cell Isolation Kit

15-Minute cell isolation kit using immunomagnetic negative selection

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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 Naïve CD4+ T Cell Isolation Kit

15-Minute cell isolation kit using immunomagnetic negative selection

From: 794 USD
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15-Minute cell isolation kit using immunomagnetic negative selection
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Product Advantages


  • Fast, easy-to-use and column-free

  • Up to 95% purity

  • Untouched, viable cells

What's Included

  • EasySep™ Mouse Naïve CD4+ T Cell Isolation Kit (Catalog #19765)
    • EasySep™ Mouse CD4+ T Cell Isolation Cocktail, 0.5 mL
    • EasySep™ Mouse Memory T Cell Depletion Cocktail, 0.5 mL
    • EasySep™ Streptavidin RapidSpheres™ 50001, 1 mL
    • EasySep™ FcR Mouse Blocker, 0.1 mL
  • RoboSep™ Mouse Naïve CD4+ T Cell Isolation Kit (Catalog #19765RF)
    • EasySep™ Mouse CD4+ T Cell Isolation Cocktail, 0.5 mL
    • EasySep™ Mouse Memory T Cell Depletion Cocktail, 0.5 mL
    • EasySep™ Streptavidin RapidSpheres™ 50001, 1 mL
    • EasySep™ FcR Mouse Blocker, 0.1 mL
    • RoboSep™ Buffer (Catalog #20104)
    • RoboSep™ Filter Tips (Catalog #20125)
Products for Your Protocol
To see all required products for your protocol, please consult the Protocols and Documentation.
  1. EasySep™ Magnet
    EasySep™ Magnet

    Magnet for column-free immunomagnetic separation

  2. EasySep™ Buffer
    EasySep™ Buffer

    Cell separation buffer

Overview

The EasySep™ Mouse Naïve CD4+ T Cell Isolation Kit is designed to isolate naïve 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-naïve CD4+ T cells (CD8, CD11b, CD11c, CD19, CD24, CD25, CD44, CD45R, CD49b, TCRγ/δ, TER119) 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.
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

Data Figures

Typical EasySep™ Mouse Naïve CD4+ T Cell Isolation Profile

Figure 1. Typical EasySep™ Mouse Naïve CD4+ T Cell Isolation Profile

Starting with mouse splenocytes from an uninfected mouse, the naïve CD4+ T cell content (CD4+CD44lowCD62Lhigh) of the isolated fraction typically ranges from 87.6 - 95.6%. In the above example, the purities of the start and final isolated fractions are 14.3% and 94.7%, respectively.

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
Document Type
Product Information Sheet 1
Product Name
RoboSep™ Mouse Naïve CD4+ T Cell Isolation Kit
Catalog #
19765RF
Lot #
1000116887 or higher
Language
English
Document Type
Product Information Sheet 2
Product Name
RoboSep™ Mouse Naïve CD4+ T Cell Isolation Kit
Catalog #
19765RF
Lot #
1000116886 or lower
Language
English
Document Type
Product Information Sheet 1
Product Name
EasySep™ Mouse Naïve CD4+ T Cell Isolation Kit
Catalog #
19765
Lot #
1000116887 or higher
Language
English
Document Type
Product Information Sheet 2
Product Name
EasySep™ Mouse Naïve CD4+ T Cell Isolation Kit
Catalog #
19765
Lot #
1000116886 or lower
Language
English
Document Type
Safety Data Sheet 1
Product Name
RoboSep™ Mouse Naïve CD4+ T Cell Isolation Kit
Catalog #
19765RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Product Name
RoboSep™ Mouse Naïve CD4+ T Cell Isolation Kit
Catalog #
19765RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 3
Product Name
RoboSep™ Mouse Naïve CD4+ T Cell Isolation Kit
Catalog #
19765RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 4
Product Name
RoboSep™ Mouse Naïve CD4+ T Cell Isolation Kit
Catalog #
19765RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 5
Product Name
RoboSep™ Mouse Naïve CD4+ T Cell Isolation Kit
Catalog #
19765RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 6
Product Name
RoboSep™ Mouse Naïve CD4+ T Cell Isolation Kit
Catalog #
19765RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Product Name
EasySep™ Mouse Naïve CD4+ T Cell Isolation Kit
Catalog #
19765
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Product Name
EasySep™ Mouse Naïve CD4+ T Cell Isolation Kit
Catalog #
19765
Lot #
All
Language
English
Document Type
Safety Data Sheet 3
Product Name
EasySep™ Mouse Naïve CD4+ T Cell Isolation Kit
Catalog #
19765
Lot #
All
Language
English
Document Type
Safety Data Sheet 4
Product Name
EasySep™ Mouse Naïve CD4+ T Cell Isolation Kit
Catalog #
19765
Lot #
All
Language
English
Document Type
Safety Data Sheet 5
Product Name
EasySep™ Mouse Naïve CD4+ T Cell Isolation Kit
Catalog #
19765
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.

Research Area
Workflow Stages

Resources and Publications

Educational Materials (9)

Tools for Your Immunology Research
Brochure
Tools for Your Immunology Research
EasySep™ Cell Separation Technology
Brochure
EasySep™ Cell Separation Technology
Mouse Cell Isolation
Brochure
Mouse Cell Isolation
Human Immune Cytokines
Wallchart
Human Immune Cytokines
Antigen Processing and Presentation
Wallchart
Antigen Processing and Presentation
How EasySep™ Magnetic Cell Separation Technology Works: Fast and Easy Cell Isolation
1:57
Video
How EasySep™ Magnetic Cell Separation Technology Works: Fast and Easy Cell Isolation
Isolate Cells with a Simple Pour-Off: EasySep™ Cell Separation Technology
1:13
Video
Isolate Cells with a Simple Pour-Off: EasySep™ Cell Separation Technology
Cell Enrichment of Mouse Naïve CD4+ T Cells from Spleen
Scientific Poster
Cell Enrichment of Mouse Naïve CD4+ T Cells from Spleen
Immunomagnetic Cell Isolation of Highly Purified Mouse Naïve CD4+ T Cells in 15 Minutes
Scientific Poster
Immunomagnetic Cell Isolation of Highly Purified Mouse Naïve CD4+ T Cells in 15 Minutes

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 (8)

Prolonged residence of an albumin-IL-4 fusion protein in secondary lymphoid organs ameliorates experimental autoimmune encephalomyelitis. A. Ishihara et al. Nature biomedical engineering 2020 oct

Abstract

Interleukin-4 (IL-4) suppresses the development of multiple sclerosis in a murine model of experimental autoimmune encephalomyelitis (EAE). Here, we show that, in mice with EAE, the accumulation and persistence in the lymph nodes and spleen of a systemically administered serum albumin (SA)-IL-4 fusion protein leads to higher efficacy in preventing disease development than the administration of wild-type IL-4 or of the clinically approved drug fingolimod. We also show that the SA-IL-4 fusion protein prevents immune-cell infiltration in the spinal cord, decreases integrin expression in antigen-specific CD4+ T cells, increases the number of granulocyte-like myeloid-derived suppressor cells (and their expression of programmed-death-ligand-1) in spinal cord-draining lymph nodes and decreases the number of T helper 17 cells, a pathogenic cell population in EAE. In mice with chronic EAE, SA-IL-4 inhibits immune-cell infiltration into the spinal cord and completely abrogates immune responses to myelin antigen in the spleen. The SA-IL-4 fusion protein may be prophylactically and therapeutically advantageous in the treatment of multiple sclerosis.
View publication
Synoviocyte-targeted therapy synergizes with TNF inhibition in arthritis reversal. M. N. D. Svensson et al. Science advances 2020 jun

Abstract

Fibroblast-like synoviocytes (FLS) are joint-lining cells that promote rheumatoid arthritis (RA) pathology. Current disease-modifying antirheumatic agents (DMARDs) operate through systemic immunosuppression. FLS-targeted approaches could potentially be combined with DMARDs to improve control of RA without increasing immunosuppression. Here, we assessed the potential of immunoglobulin-like domains 1 and 2 (Ig1{\&}2), a decoy protein that activates the receptor tyrosine phosphatase sigma (PTPRS) on FLS, for RA therapy. We report that PTPRS expression is enriched in synovial lining RA FLS and that Ig1{\&}2 reduces migration of RA but not osteoarthritis FLS. Administration of an Fc-fusion Ig1{\&}2 attenuated arthritis in mice without affecting innate or adaptive immunity. Furthermore, PTPRS was down-regulated in FLS by tumor necrosis factor (TNF) via a phosphatidylinositol 3-kinase-mediated pathway, and TNF inhibition enhanced PTPRS expression in arthritic joints. Combination of ineffective doses of TNF inhibitor and Fc-Ig1{\&}2 reversed arthritis in mice, providing an example of synergy between FLS-targeted and immunosuppressive DMARD therapies.
View publication
Mitochondrial Oxidative Phosphorylation Regulates the Fate Decision between Pathogenic Th17 and Regulatory T Cells. B. Shin et al. Cell reports 2020 feb

Abstract

Understanding metabolic pathways that regulate Th17 development is important to broaden therapeutic options for Th17-mediated autoimmunity. Here, we report a pivotal role of mitochondrial oxidative phosphorylation (OXPHOS) for lineage specification toward pathogenic Th17 differentiation. Th17 cells rapidly increase mitochondrial respiration during development, and this is necessary for metabolic reprogramming following T cell activation. Surprisingly, specific inhibition of mitochondrial ATP synthase ablates Th17 pathogenicity in a mouse model of autoimmunity by preventing Th17 pathogenic signature gene expression. Notably, cells activated under OXPHOS-inhibited Th17 conditions preferentially express Foxp3, rather than Th17 genes, and become suppressive Treg cells. Mechanistically, OXPHOS promotes the Th17 pioneer transcription factor, BATF, and facilitates T cell receptor (TCR) and mTOR signaling. Correspondingly, overexpression of BATF rescues Th17 development when ATP synthase activity is restricted. Together, our data reveal a regulatory role of mitochondrial OXPHOS in dictating the fate decision between Th17 and Treg cells by supporting early molecular events necessary for Th17 commitment.
View publication
View All Publications

Related Products

Related Products

Labeling Antibodies

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.

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.

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