EasySep™ Human CD4+ T Cell Isolation Kit

Immunomagnetic negative isolation of untouched human CD4+ T cells

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EasySep™ Human CD4+ T Cell Isolation Kit

Immunomagnetic negative isolation of untouched human CD4+ T cells

From: 964 USD
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Immunomagnetic negative isolation of untouched human CD4+ T cells
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Product Advantages


  • Fast, easy-to-use and column-free

  • Up to 97% purity with high recovery

  • Untouched, viable cells

What's Included

  • EasySep™ Human CD4+ T Cell Isolation Kit (Catalog #17952)
    • EasySep™ Human CD4+ T Cell Isolation Cocktail, 1 mL
    • EasySep™ Dextran RapidSpheres™, 1 mL
  • EasySep™ Human CD4+ T Cell Isolation (Catalog #100-0696)
    • EasySep™ Human CD4+ T Cell Isolation Cocktail, 1 x 10 mL
    • EasySep™ Dextran RapidSpheres™, 1 x 10 mL
  • RoboSep™ Human CD4+ T Cell Isolation Kit (Catalog #17952RF)
    • EasySep™ Human CD4+ T Cell Isolation Cocktail, 1 mL
    • EasySep™ Dextran RapidSpheres™, 1 mL
    • RoboSep™ Buffer (Catalog #20104)
    • RoboSep™ Filter Tips (Catalog #20125)

Overview

Easily and efficiently isolate highly purified human CD4+ T cells from fresh or previously frozen human peripheral blood mononuclear cells (PBMCs) or washed leukapheresis samples by immunomagnetic negative selection, with the EasySep™ Human CD4+ T Cell Isolation Kit. Widely used in published research for more than 20 years, EasySep™ combines the specificity of monoclonal antibodies with the simplicity of a column-free magnetic system.

In this EasySep™ negative selection procedure, unwanted cells are labeled with antibody complexes and magnetic particles. Unwanted cells expressing the following markers are targeted for removal: CD8, CD14, CD16, CD19, CD36, CD56, CD66b, CD123, TCRgd, and GlyA. The magnetically labeled cells are then separated from the untouched desired CD4+ T cells by using an EasySep™ magnet and simply pouring or pipetting the desired cells into a new tube. Following magnetic cell isolation in as little as 8 minutes, the desired CD4+ T cells are ready for downstream applications such as flow cytometry, culture, or DNA/RNA extraction.

This product replaces EasySep™ Human CD4+ T Cell Enrichment Kit (Catalog #19052) for even faster CD4+ T cell negative selections.

For large-scale isolation of human CD4+ T cells from leukapheresis samples, see the large-format (1x10^10 cells) kit (Catalog #100-0696).

Learn more about how immunomagnetic EasySep™ technology works or how to fully automate immunomagnetic cell isolation with RoboSep™. Alternatively, choose ready-to-use, ethically sourced, primary Human Peripheral Blood CD4+ T Cells, Frozen isolated with EasySep™ Human CD4+ T Cell Isolation Kit. Explore additional products optimized for your workflow, including culture media, supplements, antibodies, and more.
Magnet Compatibility
• EasySep™ Magnet (Catalog #18000)
• “The Big Easy” EasySep™ Magnet (Catalog #18001)
• Easy 50 EasySep™ Magnet (Catalog #18002)
• EasyPlate™ EasySep™ Magnet (Catalog 18102)
• EasyEights™ EasySep™ Magnet (Catalog #18103)
• RoboSep™-S (Catalog #21000)
• Easy 250 EasySep™ Magnet (Catalog #100-0821)
Subtype
Cell Isolation Kits
Cell Type
T Cells, T Cells, CD4+
Species
Human
Sample Source
Leukapheresis, PBMC
Selection Method
Negative
Application
Cell Isolation
Brand
EasySep, RoboSep
Area of Interest
Immunology

Data Figures

CD4+ T cell separation using EasySep™ Human CD4+ T Cell Isolation Kit

Figure 1. EasySep™ Human CD4+ T Cell Isolation Kit

Starting with human peripheral blood mononuclear cells (PBMCs), the CD4+ T cell (CD3+CD4+) content of the isolated fraction is typically 94.8 ± 2.3% (mean ± SD).

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 #
100-0696
Lot #
All
Language
English
Catalog #
17952
Lot #
All
Language
English
Catalog #
17952
Lot #
All
Language
Multi
Catalog #
17952RF
Lot #
All
Language
Multi
Catalog #
17952RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Catalog #
17952
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Catalog #
17952
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Catalog #
17952RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Catalog #
17952RF
Lot #
All
Language
English
Document Type
Safety Data Sheet 3
Catalog #
17952RF
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™ 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.

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.

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.

Publications (38)

Identification of Mechanisms by Which Genetic Susceptibility Loci Influence Systemic Sclerosis Risk Using Functional Genomics in Primary T Cells and Monocytes. D. Gonz\'alez-Serna et al. Arthritis & rheumatology (Hoboken, N.J.) 2023 jun

Abstract

OBJECTIVE Systemic sclerosis (SSc) is a complex autoimmune disease with a strong genetic component. However, most of the genes associated with the disease are still unknown because associated variants affect mostly noncoding intergenic elements of the genome. We used functional genomics to translate the genetic findings into a better understanding of the disease. METHODS Promoter capture Hi-C and RNA-sequencing experiments were performed in CD4+ T cells and CD14+ monocytes from 10 SSc patients and 5 healthy controls to link SSc-associated variants with their target genes, followed by differential expression and differential interaction analyses between cell types. RESULTS We linked SSc-associated loci to 39 new potential target genes and confirmed 7 previously known SSc-associated genes. We highlight novel causal genes, such as CXCR5, as the most probable candidate gene for the DDX6 locus. Some previously known SSc-associated genes, such as IRF8, STAT4, and CD247, showed cell type-specific interactions. We also identified 15 potential drug targets already in use in other similar immune-mediated diseases that could be repurposed for SSc treatment. Furthermore, we observed that interactions were directly correlated with the expression of important genes implicated in cell type-specific pathways and found evidence that chromatin conformation is associated with genotype. CONCLUSION Our study revealed potential causal genes for SSc-associated loci, some of them acting in a cell type-specific manner, suggesting novel biologic mechanisms that might mediate SSc pathogenesis.
Discovery of potent immune-modulating molecule taccaoside A against cancers from structures-active relationships of natural steroidal saponins. Z. Dai et al. Phytomedicine : international journal of phytotherapy and phytopharmacology 2022 sep

Abstract

BACKGROUND In recent years, the T-cell therapy and immune checkpoint inhibitors toward CTLA-4 and PD-1/PD-L1 axis antibody therapy have acquired encouraging success. However, most of patients were still not benefited with lots of troubles, such as low penetration of tissues/cells, strong immunogenicity and cytokine release syndrome, and long manufacturing process and expensive costs. By contrast, the immune-modulating small molecules possessed natural advantages to overcome these obstacles and might achieve greater success. PURPOSE Exploring the potent immune-modulating natural small molecules and revealing what kinds of molecules or structures with the immunomodulatory activity against cancers. METHODS A novel non-cytotoxic T-cell immunomodulating screening model was used to identify the cytotoxic/selective/immunomodulatory bioactivity for 148 natural steroidal saponins. The structure-activity relationships (SARs) research was used to reveal the key groups for immunomodulation/cytotoxicity/selectivity. The negative selection was used to isolate and purify the T-cell. The cell viability assay was used to measure the anti-cancer effect in vitro. The ELISA assay was used to detect the cytokines for IL-1$\beta$, IL-6, TNF-$\alpha$, IFN-$\gamma$, IL-12, perforin and granzyme B (GZMB). The western blotting assay was used to research the immunomodulatory mechanism. The siRNA knockdown was used to generate the IFN-$\gamma$ resistant melanoma cells. The NOG immune-deficient mice were used to evaluate the anti-tumor efficacy in vivo. The peripheral blood samples from 10 cancer patients were used to detect the broad population anti-tumor efficacy. RESULTS It was reported that the correlation among structures and immunomodulation/ cytotoxicity/selectivity, in which opening ring-F with 26-O-glucopyranosyl, disaccharide and trisaccharide chains at C-3, steric hindrance and polarity of C-22 were key immunomodulatory groups. Moreover, taccaoside A was identified as the most potent candidate against cancer cells, including non-small cell lung cancer, triple negative breast cancer, and the IFN-$\gamma$ resistant melanoma, partly through enhancing T lymphocyte mTORC1-Blimp-1 signal to secrete GZMB. Besides, 10 patients derived T-cell also would be modulated against cancer cells in vitro. Moreover, the overall survival was great extended (>140 days vs 93 days) with nearly 100% tumor burden disappearance (0 mm3vs 1006 ± 79.5 mm3) in mice. CONCLUSION This work demonstrated one possibility for this concerned purpose, and identified a potent immune-modulating natural molecule taccaoside A, which might contribute to cancer immunotherapy in future.
ABCA1, TCF7, NFATC1, PRKCZ, and PDGFA DNA methylation as potential epigenetic-sensitive targets in acute coronary syndrome via network analysis. T. Infante et al. Epigenetics 2022 may

Abstract

Acute coronary syndrome (ACS) is the most severe clinical manifestation of coronary heart disease.We performed an epigenome-wide analysis of circulating CD4+ and CD8+ T cells isolated from ACS patients and healthy subjects (HS), enrolled in the DIANA clinical trial, by reduced-representation bisulphite sequencing (RRBS). In CD4+ T cells, we identified 61 differentially methylated regions (DMRs) associated with 57 annotated genes (53% hyper- and 47% hypo-methylated) by comparing ACS patients vs HS. In CD8+ T cells, we identified 613 DMRs associated with 569 annotated genes (28% hyper- and 72% hypo-methylated) in ACS patients as compared to HS. In CD4+vs CD8+ T cells of ACS patients we identified 175 statistically significant DMRs associated with 157 annotated genes (41% hyper- and 59% hypo-methylated). From pathway analyses, we selected six differentially methylated hub genes (NFATC1, TCF7, PDGFA, PRKCB, PRKCZ, ABCA1) and assessed their expression levels by q-RT-PCR. We found an up-regulation of selected genes in ACS patients vs HS (P < 0.001). ABCA1, TCF7, PDGFA, and PRKCZ gene expression was positively associated with CK-MB serum concentrations (r = 0.75, P = 0.03; r = 0.760, P = 0.029; r = 0.72, P = 0.044; r = 0.74, P = 0.035, respectively).This pilot study is the first single-base resolution map of DNA methylome by RRBS in CD4+ and CD8+ T cells and provides specific methylation signatures to clarify the role of aberrant methylation in ACS pathogenesis, thus supporting future research for novel epigenetic-sensitive biomarkers in the prevention and early diagnosis of this pathology.
New look, same high quality and support! You may notice that your instrument or reagent packaging looks slightly different from images displayed on the website, or from previous orders. We are updating our look but rest assured, the products themselves and how you should use them have not changed. Learn more