The concentrations of cytokines and chemokines present in the culture supernatants from rhesus astrocyte and microglial cultures and rhesus FC explant cultures were quantified using the MilliPlex MAP Non-Human Primate Cytokine Magnetic Bead Panel-Premixed 23 Plex, Cytokine-Chemokine Array kit (Millipore), following the manufacturer’s instructions. The concentrations of cytokines and chemokines present in the culture supernatants from human oligodendrocytes were quantified using the Human 14-plex Cytokine-Chemokine Array kit (Millipore), following the manufacturer’s instructions. The multiplex plate was read using a Bio-Plex 200 Suspension Array Luminex System (Bio-Rad, Hercules, CA, USA).
Bio plex 200 suspension array luminex system
Manufactured by Bio-Rad
Sourced in United States
The Bio-Plex 200 Suspension Array Luminex System is a multiplex analytical platform that can simultaneously detect and quantify multiple analytes in a single sample. The system utilizes color-coded magnetic beads coated with specific capture antibodies or ligands to facilitate the measurement of various biomolecules, such as proteins, cytokines, and nucleic acids.
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Lab products found in correlation
7 protocols using bio plex 200 suspension array luminex system
1
Quantifying Cytokines and Chemokines in Cell Cultures
2
Quantifying Cytokine Profiles in Rhesus DRG and HSC
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The concentrations of cytokines and chemokines present in the culture supernatants from rhesus DRG were quantified using the MilliPlex MAP Non-Human Primate Cytokine Magnetic Bead Panel—Premixed 23 Plex, Cytokine-Chemokine Array kit (Millipore), following the manufacturer’s instructions. The analytes detected by this panel are G-CSF, GM-CSF, IFN-γ, IL-10, IL-12/23 (p40), IL-13, IL-15, IL-17, IL-18, IL-1ra, IL-1β, IL-2, IL-4, IL-5, IL-6, IL-8, CCL2, CCL3, CCL4, TGF-α, TNF, VEGF, and sCD40L. The concentrations of cytokines and chemokines present in the culture supernatants from HSC cultures described above were quantified using the Bio-Plex Pro™ Human Cytokine 27-plex Assay (BioRad, Hercules, CA). The analytes detected by this panel are FGF basic, Eotaxin, G-CSF, GM-CSF, IFN-γ, IL-1β, IL-1ra, IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-12 (p70), IL-13, IL-15, IL-17, IP-10, MCP-1 (CCL-2), MIP-1alpha, MIP-1beta, PDGF-BB, RANTES, TNF, and VEGF. The multiplex plate was read using a Bio-Plex 200 Suspension Array Luminex System (Bio-Rad).
3
Quantifying Cytokine Profiles in Macaque Lungs
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A small lung sample was excised from every lung lobe from each macaque and placed into 1mL plain DMEM (6 samples per macaque). To disrupt cell membranes, tissue samples were sonicated and then centrifuged to clarify the supernatant. Supernatants were then collected and stored at −80 °C until use. Lung homogenates were diluted at 1/4 with plain DMEM. The concentrations of cytokines and chemokines present in the lung homogenates were quantified using the MILLIPLEX MAP Non-Human Primate Cytokine Magnetic Bead Panel - PCYTMG-40 K- Cytokine-Chemokine Array Kit (Millipore) following the manufacturer’s instructions. The analytes detected by this panel are as follows: G-CSF, GM-CSF, IFN-g, IL-1ra, IL-1b, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, IL-12/23 (p40), IL-13, IL-15, IL-17, IL-18, MCP-1, MIP-1a, MIP-1b, sCD40L, TGF-a, TNF-a, and VEGF. The multiplex plate was read using a Bio-Plex 200 Suspension Array Luminex System (Bio-Rad). Data were normalized based on the tissue weight.
4
Multiplex Cytokine Profiling in CSF
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Cerebrospinal fluid (CSF) samples were collected from each animal at baseline and weeks 1, 2, 3, and 4, following infection and the concentration of cytokines and chemokines present in the (CSF) was quantified using the Bio-Plex Pro™ Human Cytokine 27-plex Assay kit (Bio-Rad, Hercules, CA) following the manufacturer’s instructions. The multiplex plate was read using a Bio-Plex 200 Suspension Array Luminex System (Bio-Rad).
5
Dexamethasone Modulates LPS-Induced IL-6 in Monocytes
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Monocytes were seeded at a density of 1x104 cells/ml in 96-well plates (Corning, Inc.) and pre-incubated with different concentrations of DEX (10-11-10-6 M) for 1 h prior to stimulation with 1 ng/ml LPS at 37˚C. After 24 h in culture, the cell supernatant was collected via centrifugation at a speed of 800 x g for 5 min at room temperature and stored at -80˚C for IL-6 analysis using the Bio-plex 200 suspension array Luminex system (Bio-rad Laboratories, Inc.). Corticosteroid sensitivity was evaluated according to the half-maximal inhibitory concentration of DEX with respect to LPS-induced IL-6 maximal production in monocytes (DEX-IC50). The percentage of maximal inhibition of IL-6 by DEX was presented as Emax. The values for individual patients were presented as log (DEX-IC50) and compared among groups.
6
Measurement of Murine Immune Markers
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Blood samples were collected by cardiac puncture 24 h after the last OVA challenge on day 35 and serum was collected by centrifugation at 2000× g for 20 min at 4 °C. Serum total IgE and OVA-specific IgG1 concentrations were measured using Enzyme-Linked Immunosorbent Assay (ELISA) kit (eBioscience, Hartfield, UK and Cayman Chemical Company, Ann Arbor, MI, USA). Concentrations of murine IL-10, IL-17, TNF-α, CXCL9, and CXCL10 in NALF were measured using Multiplex MAP kit (EMD Millipore Corp., Billerica, MA, USA) using Luminex Bio-plex 200 suspension array system (Bio-Rad Corp. Hercules, CA, USA).
7
Cytokine Profiling in Murine Asthma Model
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Concentrations of murine Th1 cytokines interferon (IFN)-γ, Th2 cytokines IL-4, IL-5, IL-13 and RANTES/CCL5, pro-inflammatory cytokine IL-17 and anti-inflammatory cytokine IL-10 in serum and BAL fluid were determined using Multiplex MAP kit (EMD Millipore Corp., Billerica, MA, USA) and Bio-plex pro assay kit reagents using Luminex Bio-plex 200 suspension array system (Bio-Rad Corp. Hercules, CA, USA). The levels of IL-31 and IL-33 in BAL fluid of the mice were measured by enzyme-linked immunosorbent assay (ELISA) kits (Thermo Fisher Scientific and BioLegend, San Diego, CA, USA, respectively). Th1/Th2 ratio was calculated based on the Th1 IFN-γ and Th2 IL-4 levels in BAL fluid. Murine OVA-specific IgE, total IgE, IL-9 and TGF-β concentrations in the serum and BAL fluid were determined using ELISA kits (BioLegend).
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