Plasma levels of soluble cytokines and adhesion molecules (IFNα, IFNγ, IL10, IL17a, IL8, MCP1, TNFα, sICAM, sVCAM) were measured prevaccination using MILLIPLEX cytokine magnetic bead panel in the MAGPIX instrument (Luminex Corporation, Austin, Texas, USA) [14 ]. MFI were analyzed with MILLIPLEX Analyst Software (EMD Millipore, Temecula, California, USA) and soluble marker levels were determined as pg/ml. sTNFR1 and II were measured using commercially available ELISA kits (R&D Systems, Minneapolis, Minnesota, USA).
Milliplex cytokine magnetic bead panel
Manufactured by DiaSorin
Sourced in United States
The MILLIPLEX cytokine magnetic bead panel is a multiplex assay designed to measure multiple cytokines in a single biological sample. It utilizes magnetic beads coated with analyte-specific capture antibodies to enable simultaneous quantification of various cytokines.
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2 protocols using milliplex cytokine magnetic bead panel
1
Quantifying Cytokine and Adhesion Profiles
2
Plasma Inflammatory and Immune Markers
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The plasma inflammatory and MT markers TNFR-I, TNFR-II and sCD14 were measured using RayBio Tech ELISA kits, (RayBiotech, Inc., GA) after appropriate dilution of the plasma according to manufacturer’s protocol. Cytokines IL-1b, IL-2, IL-6, IL-8, IL-10, IL-12 (p70), IL-17, IFN-α2, IFN-γ and TNF-α levels were determined in undiluted plasma using Milliplex cytokine magnetic bead panel in the Magpix instrument (Luminex Corporation). These markers were selected based on their known association with systemic inflammation and immune activation. Median fluorescent intensities (MFI) were analyzed and cytokine levels were expressed as pg/mL. LPS was measured in plasma samples using the Limulus amebocyte lysate chromogenic endpoint assay (Lonza, MD, USA).
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