EasySep™ Mouse CD4+CD25+ Regulatory T Cell Isolation Kit II

Immunomagnetic negative selection followed by positive selection 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.

EasySep™ Mouse CD4+CD25+ Regulatory T Cell Isolation Kit II

Immunomagnetic negative selection followed by positive selection kit

From: 952 USD
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Immunomagnetic negative selection followed by positive selection kit
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Product Advantages


  • Fast and easy-to-use

  • Up to 93% purity

  • No columns required

What's Included

  • EasySep™ Mouse CD4+CD25+ Regulatory T Cell Isolation Kit II (Catalog #18783)
    • EasySep™ Mouse CD4+ T Cell Isolation Cocktail, 0.5 mL
    • EasySep™ Streptavidin RapidSpheres™ 50001, 1 mL
    • EasySep™ Mouse CD25 Regulatory T Cell Positive Selection Cocktail, 0.5 mL
    • EasySep™ PE Selection Cocktail, 1 mL
    • EasySep™ Dextran RapidSpheres™ 50100, 1 mL
    • EasySep™ Mouse FcR Blocker (Catalog #18731), 0.5 mL
  • RoboSep™ Mouse CD4+CD25+ Regulatory T Cell Isolation Kit II (Catalog #18783RF)
    • EasySep™ Mouse CD4+ T Cell Isolation Cocktail, 0.5 mL
    • EasySep™ Streptavidin RapidSpheres™ 50001, 1 mL
    • EasySep™ Mouse CD25 Regulatory T Cell Positive Selection Cocktail, 0.5 mL
    • EasySep™ PE Selection Cocktail, 1 mL
    • EasySep™ Dextran RapidSpheres™ 50100, 1 mL
    • EasySep™ Mouse FcR Blocker (Catalog #18731), 0.5 mL
    • RoboSep™ Buffer (Catalog #20104)
    • RoboSep™ Filter Tips (Catalog #20125)
Products for Your Protocol

Overview

The EasySep™ Mouse CD4+CD25+ Regulatory T Cell Isolation Kit II is designed to isolate CD4+CD25+ T cells from single-cell suspensions of splenocytes or other tissues by negative followed by positive selection. CD4+ T cells are first enriched by negative selection using the EasySep™ Mouse CD4+ T Cell Isolation Cocktail. CD25+ cells are then positively selected from the pre-enriched cells using EasySep™ Mouse CD25 Regulatory T Cell Positive Selection Cocktail.
Magnet Compatibility
• EasySep™ Magnet (Catalog #18000)
• “The Big Easy” EasySep™ Magnet (Catalog #18001)
• EasyEights™ Magnet (Catalog #18103)
• RoboSep™-S (Catalog #21000)
Subtype
Cell Isolation Kits
Cell Type
T Cells, T Cells, Regulatory
Species
Mouse
Sample Source
Spleen
Brand
EasySep, RoboSep
Area of Interest
Immunology

Data Figures

FACS Histogram Results with EasySep™ Mouse CD4+CD25+ Regulatory T Cell Isolation Kit II

Figure 1. FACS Histogram Results with EasySep™ Mouse CD4+CD25+ Regulatory T Cell Isolation Kit II

Starting with mouse splenocytes, the regulatory T cell content (CD4+CD25+FOXP3+) of the isolated fraction typically ranges from 70 - 93%. In the above example, the purities of the start and final isolated fractions are 2% and 72%, respectively.

Protocols and Documentation

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

Document Type
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18783
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1000138480 or higher
Language
English
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18783
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1000138479 or lower
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English
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18783RF
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1000138480 or higher
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English
Catalog #
18783RF
Lot #
1000138479 or lower
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English
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Safety Data Sheet 1
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All
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English
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Safety Data Sheet 2
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Safety Data Sheet 8
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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 (4)

miR-21 and Pellino-1 Expression Profiling in Autoimmune Premature Ovarian Insufficiency. X. Li et al. Journal of immunology research 2020

Abstract

Background Premature ovarian insufficiency (POI) represents the hypergonadotropic hypoestrogenic symptoms that result in the loss of ovarian follicles. 5-30{\%} POI cases are suggested to be involved in autoimmune etiology. MicroRNA-21 (miR-21) plays a vital role in ovarian folliculogenesis via regulating and interacting with multiple target genes. Here, we conduct the target prediction of miR-21, identify the expression and correlation of miR-21 and its putative target Pellino-1 (Peli1), and confirm their relationship with clinical characteristics in autoimmune POI. Methods Bioinformatic analysis was conducted to screen the miR-21 putative target gene. Autoimmune POI mouse models were established by ZP3 immunization. Serum miR-21, Peli1 mRNA of peripheral blood mononuclear cells (PBMCs) and regulatory T cells (Tregs), general status, spleen Tregs ratio, inflammatory factors, ovarian endocrine function, and ovarian structure were evaluated. For autoimmune POI patients, serum miR-21, PBMCs Peli1 mRNA levels, general data, immune parameters, hormone levels, and ultrasound examinations were obtained. The correlations of miR-21 with Peli1 and clinical characteristics in patients were analyzed. Results Peli1 was selected based on four microRNA prediction databases and literature retrieval. In mouse models, serum miR-21 level, PBMCs and Tregs Peli1 mRNA, and spleen Tregs ratio were 0.61 ± 0.09, 0.12 ± 0.12, 0.27±0.23 and 4.82 ± 0.58, respectively, lower than those in the control group. In patients, miR-21 level (0.60 ± 0.14) and Peli1 mRNA (0.30 ± 0.14) were lower than those in the control group (1.01 ± 0.07 and 1.63 ± 0.54); miR-21 was positively related with Peli1, AMH, E2, the size of the uterus, and ovarian volume and negatively related with FSH, LH, and the number of positive immune parameters (AOAb, EMAb, ACL, ANA, ds-DNA, ACA, IgG, IgA, IgM, IgE, C3, and C4). Conclusions Low expressions of miR-21 and Peli1 were detected in autoimmune POI mice and patients. Positive correlation between miR-21 and Peli1 was observed in autoimmune POI patients, suggesting that miR-21 and Peli1 might be associated with the pathogenesis of autoimmune POI.
IL233, an IL-2-IL-33 hybrid cytokine induces prolonged remission of mouse lupus nephritis by targeting Treg cells as a single therapeutic agent. M. E. Stremska et al. Journal of autoimmunity 2019 may

Abstract

Lupus glomerulonephritis (GN) is an autoimmune disease characterized by immune complex-deposition, complement activation and glomerular inflammation. In lupus-prone NZM2328 mice, the occurrence of lupus GN was accompanied by a decrease in Treg cells and an increase in proinflammatory cytokine-producing T cells. Because IL-33 in addition to IL-2 has been shown to be important for Treg cell proliferation and ST2 (IL-33 receptor) positive Treg cells are more potent in suppressor activity, a hybrid cytokine with active domains of IL-2 and IL-33 was generated to target the ST2+ Treg cells as a therapeutic agent to treat lupus GN. Three mouse models were used: spontaneous and Ad-IFNalpha- accelerated lupus GN in NZM2328 and the lymphoproliferative autoimmune GN in MRL/lpr mice. Daily injections of IL233 for 5 days prevented Ad-IFNalpha-induced lupus GN and induced remission of spontaneous lupus GN. The remission was permanent in that no relapses were detected. The remission was accompanied by persistent elevation of Treg cells in the renal lymph nodes. IL233 is more potent than IL-2 and IL-33 either singly or in combination in the treatment of lupus GN. The results of this study support the thesis that IL233 should be considered as a novel agent for treating lupus GN.
Nanoparticles Containing an Insulin-ChgA Hybrid Peptide Protect from Transfer of Autoimmune Diabetes by Shifting the Balance between Effector T Cells and Regulatory T Cells. B. L. Jamison et al. Journal of immunology (Baltimore, Md. : 1950) 2019 jul

Abstract

CD4 T cells play a critical role in promoting the development of autoimmunity in type 1 diabetes. The diabetogenic CD4 T cell clone BDC-2.5, originally isolated from a NOD mouse, has been widely used to study the contribution of autoreactive CD4 T cells and relevant Ags to autoimmune diabetes. Recent work from our laboratory has shown that the Ag for BDC-2.5 T cells is a hybrid insulin peptide (2.5HIP) consisting of an insulin C-peptide fragment fused to a peptide from chromogranin A (ChgA) and that endogenous 2.5HIP-reactive T cells are major contributors to autoimmune pathology in NOD mice. The objective of this study was to determine if poly(lactide-co-glycolide) (PLG) nanoparticles (NPs) loaded with the 2.5HIP Ag (2.5HIP-coupled PLG NPs) can tolerize BDC-2.5 T cells. Infusion of 2.5HIP-coupled PLG NPs was found to prevent diabetes in an adoptive transfer model by impairing the ability of BDC-2.5 T cells to produce proinflammatory cytokines through induction of anergy, leading to an increase in the ratio of Foxp3+ regulatory T cells to IFN-gamma+ effector T cells. To our knowledge, this work is the first to use a hybrid insulin peptide, or any neoepitope, to re-educate diabetogenic T cells and may have significant implications for the development of an Ag-specific therapy for type 1 diabetes patients.
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.