EasySep™ Mouse CD4+CD62L+ T Cell Isolation Kit

Immunomagnetic negative selection followed by positive selection kit

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Immunomagnetic negative selection followed by positive selection kit
From: 751 USD

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Overview

Isolate highly purified naïve CD4+ T cells from single-cell suspensions of splenocytes or other tissues by using a simple, two-step method. First, naive CD4+ T cells are pre-enriched by negative selection using the EasySep™ Mouse Naïve CD4+ T Cell Pre-Enrichment Cocktail containing biotinylated antibodies directed against non-naïve CD4+ T cells (CD8, CD11b, CD11c, CD19, CD24, CD25, CD45R, CD49b, TCRγ/δ, TER119). Then CD4+CD62L+ cells, pre-labeled with CD62L PE, are selected using the EasySep™ PE Selection Cocktail. The desired cells are labeled with antibodies and magnetic particles. The cells are separated without columns using an EasySep™ magnet.

For the isolation of mouse naïve CD4+ T cells in as little as 15 minutes by negative selection, we recommend using the EasySep™ Mouse Naïve CD4+ T Cell Isolation Kit (19765).
Advantages:
• Fast and easy-to-use
• Up to 97% purity
• No columns required
Components:
  • EasySep™ Mouse CD4+CD62L+ T Cell Isolation Kit (Catalog #18765)
    • EasySep™ Mouse Naïve CD4+ T Cell Pre-Enrichment Cocktail, 0.5 mL
    • EasySep™ Streptavidin RapidSpheres™ 50001, 1 mL
    • EasySep™ PE Positive Selection Cocktail, 1 mL
    • EasySep™ Dextran RapidSpheres™ 50100, 1 mL
    • Normal Rat Serum, 2 mL
  • RoboSep™ Mouse CD4+CD62L+ T Cell Isolation Kit (Catalog #18765RF)
    • EasySep™ Mouse Naïve CD4+ T Cell Pre-Enrichment Cocktail, 0.5 mL
    • EasySep™ Streptavidin RapidSpheres™ 50001, 1 mL
    • EasySep™ PE Positive Selection Cocktail, 1 mL
    • EasySep™ Dextran RapidSpheres™ 50100, 1 mL
    • Normal Rat Serum, 2 mL
    • RoboSep™ Buffer (Catalog #20104)
    • RoboSep™ Filter Tips (Catalog #20125) x 2
Magnet Compatibility:
• EasySep™ Magnet (Catalog #18000)
• “The Big Easy” EasySep™ Magnet (Catalog #18001)
• RoboSep™-S (Catalog #21000)
Subtype:
Cell Isolation Kits
Cell Type:
T Cells; T Cells, CD4+
Species:
Mouse
Sample Source:
Other; Spleen
Selection Method:
Positive
Application:
Cell Isolation
Brand:
EasySep; RoboSep
Area of Interest:
Immunology

Scientific Resources

Educational Materials

(9)

Product 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.

Data and Publications

Data

Typical EasySep™ Mouse CD4+CD62L+ T Cell Isolation Profile

Figure 1. Typical EasySep™ Mouse CD4+CD62L+ T Cell Isolation Profile

Starting with a single-cell suspension of splenocytes, the naïve CD4 + T cell (CD4+CD44low CD62Lhigh) content of the final isolated fraction typically ranges from 91.7% - 96.7%. In the example above, the final purities of the start and isolated fractions are 17.9% and 92.4%, respectively.

Publications

(1)
Journal of autoimmunity 2019 may

NF-kappaB-driven miR-34a impairs Treg/Th17 balance via targeting Foxp3.

M. Xie et al.

Abstract

The subset of regulatory T (Treg) cells, with its specific transcription Foxp3, is a unique cell type for the maintenance of immune homeostasis by controlling effector T (Teff) cell responses. Although it is common that a defect in Treg cells with Treg/Teff disorder causes autoimmune diseases; however, the precise mechanisms are not thoroughly revealed. Here, we report that miR-34a could attenuate human and murine Foxp3 gene expression via targeting their 3' untranslated regions (3' UTR). The human miR-34a, increased in peripheral blood mononuclear cells (PBMCs) and CD4+ T cells from rheumatoid arthritis (RA) or systemic lupus erythematosus (SLE) patients, displayed a positive correlation with some serum markers of inflammation including rheumatoid factor (RF), anti-streptolysin antibody (ASO), erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP) as well as Th17 signature gene RORgammat, but inversely correlated with the mRNA expression levels of FOXP3. In addition, murine miR-34a levels were downregulated in TGF-beta-induced Treg cells but upregulated in Th17 cells induced in vitro compared to activated CD4+ T cells. It has also been demonstrated that elevated miR-34a disrupting Treg/Th17 balance in vivo contributed to the progress of pathogenesis of collagen induced arthritis (CIA) mice. Furthermore, IL-6 and TNF-alpha were responsible for the upregulation of miR-34a and downregulation of Foxp3, which was reverted by the addition of NF-kappaB/p65 inhibitor BAY11-7082, thus indicating that NF-kappaB/p65 inhibited Foxp3 expression in an miR-34a-dependent manner. Finally, IL-6 or TNF-alpha-activated p65 could bind to the miR-34a promotor and enhance its activity, resulting in upregulation of its transcription. Taken together, we show that NF-kappaB activated by inflammatory cytokines, such as IL-6 and TNF-alpha, ameliorates Foxp3 levels via regulating miR-34a expression, which provides a new mechanistic and therapeutic insight into the ongoing of autoimmune diseases.
STEMCELL TECHNOLOGIES INC.’S QUALITY MANAGEMENT SYSTEM IS CERTIFIED TO ISO 13485. PRODUCTS ARE FOR RESEARCH USE ONLY AND NOT INTENDED FOR HUMAN OR ANIMAL DIAGNOSTIC OR THERAPEUTIC USES UNLESS OTHERWISE STATED.