From venous whole blood collected from all patients, T-regulatory (T-reg) lymphocytes were isolated. The principle of isolation of CD4/CD25 cells from blood is based on the protocol of two-step selection of T-regulatory lymphocytes (T-reg), according to Sugiyama H. et al., 2005 [41 (link)] (with minor modifications and the use of kits StemCell Technologies, Vancouver, BC, Kanada). Firstly, CD4+ cells were obtained by negative selection using the RosetteSep®Humane CD4+ T Cell Enrichment Cocktail kit (StemCell Technologies), according to the manufacturer’s instructions. Following this, CD25+ T lymphocytes were positively selected from enriched CD4+ cells using the EasySep® Human CD25 Positive Selection Cocktail kit (StemCell Technologies) and magnetic beads EasySep® Magnetic Nanoparticles (StemCell Technologies). The positively-selected cells were resuspended in 1 × PBS and used for RNA isolation.
Easysep human cd25 positive selection cocktail kit
Manufactured by STEMCELL
Sourced in Canada
The EasySep® Human CD25 Positive Selection Cocktail kit is a laboratory product designed to isolate CD25-positive cells from human samples. The kit contains a cocktail of antibodies and magnetic particles that specifically target and bind to CD25-expressing cells, allowing their separation from the rest of the cell population using a compatible magnetic cell separation system.
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Lab products found in correlation
2 protocols using easysep human cd25 positive selection cocktail kit
1
Isolation of Human CD4+CD25+ T-Regulatory Cells
2
Two-Stage T-Reg Cell Isolation from Blood
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In the venous whole blood collected from all patients, 2-stage selection of T-regulatory lymphocytes was performed. The first step was obtaining CD4+ cells using the RosetteSep ® Humane CD4+ T Cell Enrichment Coctail kit (StemCell Technologies, Vancouver, Canada), according to the manufacturer's protocol. In the second stage, CD25+ T lymphocytes were selected from enriched CD4+ cells using the EasySep ® Human CD25 Positive Selection Cocktail kit (StemCell Technologies, Vancouver, Canada) and magnetic beads EasySep ® Magnetic Nanoparticles (StemCell Technologies, Vancouver, Canada). The positively selected cells were resuspended in 1 × PBS and destined for RNA isolation.
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