EasySep™ Human Resting CD4+ T Cell Isolation Kit

Immunomagnetic negative selection kit
概要
The EasySep™ Human Resting CD4+ T Cell Isolation Kit is designed to isolate resting CD4+ T cells from fresh or previously frozen human peripheral blood mononuclear cells or washed leukapheresis samples by immunomagnetic negative selection. The EasySep™ procedure involves labeling unwanted cells with antibody complexes and magnetic particles. The magnetically labeled cells are separated from the untouched desired cells by using an EasySep™ magnet and simply pouring or pipetting the desired cells into a new tube.
Advantages
• Fast, easy-to-use and column-free
• Up to 99% purity
• Isolated cells are untouched
Components
  • EasySep™ Human Resting CD4+ T Cell Isolation Kit (Catalog #17962)
    • EasySep™ Human Resting CD4+ T Cell Isolation Cocktail, 1 mL
    • EasySep™ Human CD25 Depletion Cocktail, 1 mL
    • EasySep™ Dextran RapidSpheres™, 2 x 1 mL
    • RoboSep™ Empty Vial
  • RoboSep™ Human Resting CD4+ T Cell Isolation Kit (Catalog #17962RF)
    • EasySep™ Human Resting CD4+ T Cell Isolation Cocktail, 1 mL
    • EasySep™ Human CD25 Depletion Cocktail, 1 mL
    • EasySep™ Dextran RapidSpheres™, 2 x 1 mL
    • RoboSep™ Empty Vial
    • RoboSep™ Buffer (Catalog #20104)
    • RoboSep™ Filter Tips (Catalog #20125)
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
Human
Sample Source
Leukapheresis, PBMC
Selection Method
Negative
Application
Cell Isolation
Brand
EasySep, RoboSep
Area of Interest
Immunology
数据及文献

Data

(A) Starting with fresh PBMCs, the resting CD4+ T cell content (CD3+CD4+CD8-CD25-CD69-HLA-DR-) of the isolated fraction is typically 95.5 ± 6.5% (mean ± SD using the purple EasySep™ Magnet). In the above example, the purities of the start and final isolated fractions are 35% and 99%, respectively. (B) Starting with fresh PBMCs that have elevated CD25/CD69/HLA-DR expression, the resting CD4+ T cell content of the isolated fraction is typically 88.0 ± 7.0% (mean ± SD using the purple EasySep™ Magnet). In the above example, the purities of the start and final isolated fractions are 8% and 85%, respectively.

Publications (1)

Journal of clinical medicine 2020 jul Hepatitis C Virus Influences HIV-1 Viral Splicing in Coinfected Patients. P. Mart\'inez-Rom\'an et al.

Abstract

Coinfection with hepatitis C virus (HCV) influences HIV reservoir size. However, it is unknown whether this coinfection also induces a higher provirus transcription. Viral transcription is promoted by synergy between cellular factors such as NF-$\kappa$B and the viral regulator Tat. The impact of HCV coinfection on HIV provirus transcription was analyzed in resting (r)CD4 T+ cells (CD3+CD4+CD25-CD69-HLADR-) and rCD4 T cells-depleted PBMCs (rCD4 T- PBMCs) from a multicenter cross-sectional study of 115 cART-treated HIV patients: 42 HIV+/HCV+ coinfected individuals (HIV+/HCV+), 34 HIV+ patients with HCV spontaneous clearance (HIV+/HCV-) and 39 HIV patients (HIV+). Viral transcription was assessed in total RNA through the quantification of unspliced, single spliced, and multiple spliced viral mRNAs by qPCR. Linear correlations between viral reservoir size and viral splicing were determined. A 3-fold increase of multiple spliced transcripts in rCD4 T+ cells of HIV+/HCV+ patients was found compared to HIV+ individuals (p {\textless} 0.05). As Tat is synthesized by multiple splicing, the levels of Tat were also quantified in these patients. Significant differences in single and multiple spliced transcripts were also observed in rCD4 T- PBMCs. Levels of multiple spliced mRNAs were increased in rCD4 T+ cells isolated from HIV+/HCV+ subjects, which could indicate a higher Tat activity in these cells despite their resting state.
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