After dilution to 1:5, nasal fluids were analysed for the concentration of various, below-mentioned cytokines using a human cytokine 27-plex panel according to manufacturer`s instructions (Bio-Plex Pro Human Cytokine Standard 27-Plex, Group I, Bio-Rad Laboratories, Hercules, California, USA). This assay uses fluorescently labelled polystyrene beads which are conjugated to capture antibodies directed to the cytokines. After washing, the fluorescently addressed detection antibody forms an immunoassay with the cytokine. For analysis, the fluorochromes are excited by two lasers: one for classifying each bead, the other for quantifying the amount of bound substrate [25 (link)]. The same procedure was performed with plasma, however, after dilution to 1:4. The detection threshold was 0.5 pg/ml. The determined cytokines were eotaxin as eosinophilic marker, IL-4, IL-5 and IL-13 as part of the Th2-cluster, IL2, IL-12 and IFN-gamma as markers of the Th1-cluster, IL-10 as representative of the Treg-Cluster, and unspecific inflammatory markers such as IL-1β, IL-2, IL-6, IL-7, IL-8, IL-17, TNF-alpha, MCP-1, MIP-1β and IP-10.
Bio plex pro human cytokine standard 27 plex
Manufactured by Bio-Rad
Sourced in United States
The Bio-Plex Pro Human Cytokine Standard 27-plex is a laboratory equipment product offered by Bio-Rad. It is a pre-mixed panel of 27 different human cytokines and chemokines designed for use with the Bio-Plex suspension array system.
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3 protocols using bio plex pro human cytokine standard 27 plex
1
Nasal Fluid Cytokine Profiling in Research
2
Multiplex Cytokine Quantification
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Quantification of cytokine levels from culture supernatants was performed using a human 27-plex assay (Biorad Bio-Plex Pro Human Cytokine Standard 27-plex, Group I), on the Bioplex 200 (Biorad, California, USA). The panel comprised of Interleukin (IL)-1β, IL-1ra, IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL12p70, IL-13, IL-15, IL17A, Eotaxin, Basic Fibroblast growth factor (FGF), Granulocyte-Colony Stimulating Factor (G-CSF), Granulocyte-Macrophage Colony Stimulating Factor (GM-CSF), Interferon gamma (IFNγ), Interferon Gamma-Induced Protein (IP)-10, Monocyte chemoattractant protein (MCP)-1, Macrophage Inflammatory Protein (MIP)-1α, MIP-1β, Platelet-derived growth factor (PDGF), Regulated on Activation, Normal T-cell Expressed and Secreted (RANTES), Tumour Necrosis Factor alpha (TNFα) and Vascular Endothelial Growth Factor (VEGF). Measurements were examined using Bio-Plex Manager v.4.1.1. software against internal quality controls and dilutions of cytokine standard curves were prepared according to manufacturer’s instructions.
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Multiplex Cytokine Quantification
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