To determine systemic inflammation, plasma concentrations of G-CSF, IFNγ, IL-1α, IL-1β, IL-2, IL-3, IL-9, IL-12(p20) IL-13, IL-15, LIF, LIX, MCP-1, M-CSF, MIG, MIP-2, RANTES, and TNF-α were measured using Mouse Cytokine/Chemokine Magnetic Bead Panel (Milliplex, St. Louis, MO) following manufacturers protocol for plasma samples. Samples were not further diluted following pretreatment when HemogloBind (dilution factor following treatment of 1.75). Samples were run on Luminex FLEXMAP 3D® instrument system with xPONENT software (EMD Millipore; St. Louis, MO) following the manufacturer's recommendation. Median fluorescent intensity data was used to determine unknown concentrations in Milliplex Analyst Software (EMD Millipore, St. Louis, MO). Samples measuring outside of the standard curve were removed from the analysis (n = 3–6). For supplementary data n = 2–8 and groups with all measurements below the standard curve are represented as no data (ND).
Flexmap 3d
Manufactured by Merck Group
Sourced in United States
The FLEXMAP 3D is a multiplex assay system designed for high-throughput analysis. It utilizes color-coded magnetic beads and laser-based detection to enable simultaneous measurement of multiple analytes in a single sample. The system is capable of analyzing up to 500 different analytes per sample.
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Lab products found in correlation
3 protocols using flexmap 3d
1
Multiplex Analysis of Inflammation Biomarkers
2
Comprehensive Plasma Complement Analysis
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Plasma study samples were also assessed for complement using the Merck Millipore Milliplex Human Complement Panel 1 (HCMP1MAG‐19K) and Human Complement Panel 2 (HCMP2MAG19K) (Merck Millipore, Massachusetts, USA), following the manufacturer’s recommendations on a Luminex FlexMap 3D. The plasma samples were prepared at a dilution of 1:200 as recommended for Panel 1 and 1:40 000 as recommended by Panel 2. The following analytes were assessed in Panel 1: C2, C4b, C5, C9, Factor D (Adipsin), Mannose Binding Lectin (MBL) and Factor I. The following analytes were assessed in Panel 2: C1q, C3, C3b/iC3b, C4, Factor B, Factor H and Properdin.
3
Multiplex Cytokine Profiling of Post-Vaccination Plasma
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A Milliplex© human cytokine/chemokine panel (Cat: EPX090-12187-901; Merck Millipore, Massachusetts, USA) was used to measure plasma levels of immunological molecules on days 0, 2, 7, and 28 post-vaccination. Samples and standards were incubated with fluorescent-coded magnetic beads that had been precoated with the respective capture antibodies. After overnight incubation at 4°C with shaking, the plates were washed twice with the provided wash buffer. Biotinylated detection antibodies were incubated with the complex for 1 h and Streptavidin-PE was subsequently added for a further 30-min incubation. The plates were washed twice before the beads were re-suspended in sheath fluid and read as PCR plates by the Luminex analyzer, FLEXMAP© 3D (Merck Millipore, Massachusetts, USA). The data were acquired using xPONENT© 4.0 (Luminex©, Texas, USA) acquisition software and analyzed with Bio-Plex Manager 6.1.1© (Bio-Rad, California, USA). Standard curves were generated, and a five-parameter logistic algorithm was used to estimate the MFI and concentration values.
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