Isolated RNA was processed (1 μg) with reverse transcriptase (Qiagen). Gene‐specific PCR products were measured by CFX96 TouchTM Real‐Time PCR Detection System (Bio‐Rad, Munich, Germany) for 40 cycles. Subpopulations of 2.5 × 106 PBMC were counted on a FACSCanto II flow cytometer (Becton Dickinson). Cytokine levels were determined in supernatants of cultured PBMCs with Human Cytokine Multiplex Assay Kit (Bio‐Rad) using LUMINEX.
Human cytokine multiplex assay kit
Manufactured by Bio-Rad
Sourced in Germany
The Human Cytokine Multiplex Assay Kit is a lab equipment product from Bio-Rad. It is designed to quantitatively measure multiple human cytokines, chemokines, and growth factors in a single sample.
Automatically generated - may contain errors
Lab products found in correlation
Reverse transcriptase, by Qiagen (1 mentions)
Cfx96 touch real time pcr detection system, by Bio-Rad (1 mentions)
Facscanto 2, by BD (1 mentions)
Human serum, by Thermo Fisher Scientific (1 mentions)
Rpmi 1640, by Thermo Fisher Scientific (1 mentions)
Ficoll paque plus, by GE Healthcare (1 mentions)
Phosphate buffered saline (pbs), by Thermo Fisher Scientific (1 mentions)
2 protocols using human cytokine multiplex assay kit
1
Profiling Immune Cell Subsets and Cytokines
2
Cytokine Profiles of Neonatal Immune Cells
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Cord blood (20 ml) and peripheral blood (5–10 ml) samples were collected from healthy neonates within 24 h of birth. Cord blood mononuclear cells (CBMCs) were isolated from cord blood by density gradient centrifugation with Ficoll-Paque PLUS (GE Healthcare, Piscataway, NJ, USA) after dilution in PBS (Gibco, Carlsbad, CA, USA). Cells were washed in RPMI 1640 (Gibco) and diluted in 10 % human serum (Gibco) to a concentration of 5 × 106 cells/ml. CBMCs were cultured for three days in the presence of 10 μg/ml PPG, 0.1 μg/ml LPA (Sigma-Aldrich, St. Louis, MO, USA), or without a stimulation factor. Concentrations of IL-10, IL-13, IL-17, and interferon-γ (IFN-γ) in the CBMC supernatants were measured with a Human Cytokine Multiplex Assay kit (Bio-Rad, Munich, Germany) with Luminex detection. The lower limits of detection for the assay (pg/mL) were: 0.9 (IL-10), 2.1 (IL-13), 0.2 (IL-17), and 1.3 (IFN-γ).
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