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EasySep™ Mouse Memory CD4+ T Cell Isolation Kit

27.5-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 Memory CD4+ T Cell Isolation Kit

27.5-Minute cell isolation kit using immunomagnetic negative selection

From: 791 USD
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27.5-Minute cell isolation kit using immunomagnetic negative selection
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Product Advantages


  • Fast, easy-to-use and column-free

  • Up to 96% purity

  • Untouched, viable cells

What's Included

  • EasySep™ Mouse Memory CD4+ T Cell Isolation Kit (Catalog #19767)
    • EasySep™ Mouse Memory CD4+ T Cell Isolation Cocktail, 0.5 mL
    • EasySep™ Streptavidin RapidSpheres™ 50001, 2 x 1 mL
    • EasySep™ Mouse FcR Blocker (Catalog #18730), 0.2 mL
  • RoboSep™ Mouse Memory CD4+ T Cell Isolation Kit (Catalog #19767RF)
    • EasySep™ Mouse Memory CD4+ T Cell Isolation Cocktail, 0.5 mL
    • EasySep™ Streptavidin RapidSpheres™ 50001, 2 x 1 mL
    • EasySep™ Mouse FcR Blocker (Catalog #18730), 0.2 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. EasyEights™ EasySep™ Magnet
    EasyEights™ EasySep™ Magnet

    Multiple sample processing magnet for column-free immunomagnetic cell separation

  2. EasySep™ Buffer
    EasySep™ Buffer

    Cell separation buffer

Overview

The EasySep™ Mouse Memory CD4+ T Cell Isolation Kit is designed to isolate pan-memory 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-memory CD4+ T cells (CD8, CD11b, CD11c, CD19, CD24, CD45R, CD45RB, CD49b, CD105 and TCRγ/δ) 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 Memory CD4+ T Cell Isolation Profile

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

Starting with mouse splenocytes, the memory CD4+ T cell (CD4+CD62L high CD44 high/int central memory and CD4+CD62L-CD44 high/int effector memory) content of the isolated fraction typically ranges from 78 - 96%. In the example above, the final purities of the start and isolated fractions are 5.3% and 90.2%, 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 Memory CD4+ T Cell Isolation Kit
Catalog #
19767RF
Lot #
1000157900 or higher
Language
English
Document Type
Product Information Sheet 2
Product Name
RoboSep™ Mouse Memory CD4+ T Cell Isolation Kit
Catalog #
19767RF
Lot #
1000157899 or lower
Language
English
Document Type
Product Information Sheet 1
Product Name
EasySep™ Mouse Memory CD4+ T Cell Isolation Kit
Catalog #
19767
Lot #
1000157899 or lower
Language
English
Document Type
Product Information Sheet 2
Product Name
EasySep™ Mouse Memory CD4+ T Cell Isolation Kit
Catalog #
19767
Lot #
1000157900 or higher
Language
English
Document Type
Safety Data Sheet 1
Product Name
RoboSep™ Mouse Memory CD4+ T Cell Isolation Kit
Catalog #
19767RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Product Name
RoboSep™ Mouse Memory CD4+ T Cell Isolation Kit
Catalog #
19767RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 3
Product Name
RoboSep™ Mouse Memory CD4+ T Cell Isolation Kit
Catalog #
19767RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 4
Product Name
RoboSep™ Mouse Memory CD4+ T Cell Isolation Kit
Catalog #
19767RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 5
Product Name
RoboSep™ Mouse Memory CD4+ T Cell Isolation Kit
Catalog #
19767RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Product Name
EasySep™ Mouse Memory CD4+ T Cell Isolation Kit
Catalog #
19767
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Product Name
EasySep™ Mouse Memory CD4+ T Cell Isolation Kit
Catalog #
19767
Lot #
All
Language
English
Document Type
Safety Data Sheet 3
Product Name
EasySep™ Mouse Memory CD4+ T Cell Isolation Kit
Catalog #
19767
Lot #
All
Language
English
Document Type
Safety Data Sheet 4
Product Name
EasySep™ Mouse Memory CD4+ T Cell Isolation Kit
Catalog #
19767
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)

EasySep™ Cell Separation Technology
Brochure
EasySep™ Cell Separation Technology
Tools for Your Immunology Research
Brochure
Tools for Your Immunology Research
Mouse Cell Isolation
Brochure
Mouse Cell Isolation
Antigen Processing and Presentation
Wallchart
Antigen Processing and Presentation
Human Immune Cytokines
Wallchart
Human Immune Cytokines
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
Simultaneous Cell Isolation from Multiple Samples Using the EasyEights™ EasySep™ Magnet
0:57
Video
Simultaneous Cell Isolation from Multiple Samples Using the EasyEights™ EasySep™ Magnet
Streamline Your Mouse T Cell Research from Isolation to Expansion
30:18
Webinar
Streamline Your Mouse T Cell Research from Isolation to Expansion

Frequently Asked Questions

Can EasySep™ be used for either positive or negative selection?

Yes. The EasySep™ kits use either a negative selection approach by targeting and removing unwanted cells or a positive selection approach targeting desired cells. Depletion kits are also available for the removal of cells with a specific undesired marker (e.g. GlyA).

How does the separation work?

Magnetic particles are crosslinked to cells using Tetrameric Antibody Complexes (TAC). When placed in the EasySep™ Magnet, labeled cells migrate to the wall of the tube. The unlabeled cells are then poured off into a separate fraction.

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™ separations be automated?

Yes. RoboSep™, the fully automated cell separator, automates all EasySep™ labeling and cell separation steps.

Can EasySep™ be used to isolate rare cells?

Yes. We recommend a cell concentration of 2x108 cells/mL and a minimum working volume of 100 µL. Samples containing 2x107 cells or fewer should be suspended in 100 µL of buffer.

Are the EasySep™ magnetic particles FACS-compatible?

Yes, the EasySep™ particles are flow cytometry-compatible, as they are very uniform in size and about 5000X smaller than other commercially available magnetic beads used with column-free systems.

Can the EasySep™ magnetic particles be removed after enrichment?

No, but due to the small size of these particles, they will not interfere with downstream applications.

How does the binding of the EasySep™ magnetic particle affect the cells? is the function of positively selected cells altered by the bound particles?

Hundreds of publications have used cells selected with EasySep™ positive selection kits for functional studies. Our in-house experiments also confirm that selected cells are not functionally altered by the EasySep™ magnetic particles.

If particle binding is a key concern, we offer two options for negative selection. The EasySep™ negative selection kits can isolate untouched cells with comparable purities, while RosetteSep™ can isolate untouched cells directly from whole blood without using particles or magnets.

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.

For positive selection, can I perform more than 3 separations to increase purity?

Yes, the purity of targeted cells will increase with additional rounds of separations; however, cell recovery will decrease.

Which cell separation kits are compatible with the EasyPlate™ EasySep™ magnet?

The EasyPlate™-compatible human EasySep™ kits are:
19051 (T Cells), 19052 (CD4 T cells), 19157 (Memory CD4 T Cells), 19053 (CD8 T Cells), 19054 (B Cells), 19254 (Naïve B cells), 19055 (NK Cells), 19058 (Monocytes without CD16 depletion), 19059 (Monocytes)

The EasyPlate™-compatible mouse EasySep™ kits are:
19751 (T Cells), 19752 (CD4 T cells), 19753 (CD8 T Cells), 19754 (B Cells), 19755 (NK Cells - please contact Tech Support)

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 (1)

T cell-intrinsic role for Nod2 in protection against Th17-mediated uveitis. R. J. Napier et al. Nature communications 2020

Abstract

Mutations in nucleotide-binding oligomerization domain-containing protein 2 (NOD2) cause Blau syndrome, an inflammatory disorder characterized by uveitis. The antimicrobial functions of Nod2 are well-established, yet the cellular mechanisms by which dysregulated Nod2 causes uveitis remain unknown. Here, we report a non-conventional, T cell-intrinsic function for Nod2 in suppression of Th17 immunity and experimental uveitis. Reconstitution of lymphopenic hosts with Nod2-/- CD4+ T cells or retina-specific autoreactive CD4+ T cells lacking Nod2 reveals a T cell-autonomous, Rip2-independent mechanism for Nod2 in uveitis. In naive animals, Nod2 operates downstream of TCR ligation to suppress activation of memory CD4+ T cells that associate with an autoreactive-like profile involving IL-17 and Ccr7. Interestingly, CD4+ T cells from two Blau syndrome patients show elevated IL-17 and increased CCR7. Our data define Nod2 as a T cell-intrinsic rheostat of Th17 immunity, and open new avenues for T cell-based therapies for Nod2-associated disorders such as Blau syndrome.
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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