Protein lysates were generated from 1-cm proximal colon samples as described previously (Zhou et al., 2023 (link)). PB was obtained by cardiac puncture and allowed to clot for 1-2 hours at room temperature. Following centrifugation at 2000 rpm for 10 minutes (4°C), serum was removed and stored at −80°C until use. Colon lysates and serum samples were subjected to a bead-based multiplex assay (Cytokine & Chemokine 36-Plex Mouse ProcartaPlex Panel 1A, Invitrogen, Waltham, MA) performed on a Luminex 200 platform according to the manufacturer’s instructions. A 96-well plate assay was designed to analyze an array of 36 cytokines and chemokines (CCL2, CCL3, CCL4, CCL5, CCL7, CCL11, CXCL1, CXCL2, CXCL5, CXCL10, G-CSF, GM-CSF, IFN-α, IFN-γ, IL-1α, IL1-β, IL-2, IL-3, IL-4, IL-5, IL-6, IL-9, IL-10, IL12p70, IL-13, IL-15, IL-17A, IL-22, IL-23, IL-27, IL-28, IL-29, IL-31, LIF, M-CSF, and TNF-α). All samples were assayed in triplicate, and standards were measured in duplicate.
Luminex 200 platform
Manufactured by Thermo Fisher Scientific
The Luminex 200 platform is a multiplex analytical system designed for the simultaneous detection and quantification of multiple analytes in a single sample. It utilizes color-coded magnetic microspheres, flow cytometry, and proprietary xMAP technology to perform various types of immunoassays and molecular diagnostics.
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Lab products found in correlation
2 protocols using luminex 200 platform
1
Multiplex Cytokine Profiling of Colon and Serum
2
Plasma Cytokine Quantification Protocol
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Retro-orbital sinus blood was collected on post-injection days 7 and 24. Blood samples were centrifuged at 1000× g at 4 °C for 10 min to acquire plasma fractions. Plasma IFN-β (EPX01B-20644-901, Invitrogen™, Carlsbad, CA, USA), IFN-γ (EPX01A-20606-901, Invitrogen™), and TNF-α (EPXS010-20607-901, Invitrogen™) were measured according to the manufacturer’s instructions (MAN0016941, ProcartaPlex™ Multiplex Immunoassay, Invitrogen™). Briefly, plasma cytokine levels were measured using primary antibodies conjugated to magnetic beads and quantified on a Luminex™ 200 platform (Invitrogen™).
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