Sera were collected from the blood of all mice on day 45; the content of cytokines (TNF-a, IL-6, and IL-10) was measured by using the mouse magnetic Luminex screening assay (R&D Systems, Minneapolis, MN, USA).
Mouse magnetic luminex screening assay
Manufactured by R&D Systems
Sourced in United States
The Mouse Magnetic Luminex Screening Assay is a multiplex assay designed to measure multiple analytes in a single sample. It utilizes magnetic beads coated with specific capture antibodies to detect and quantify various proteins in mouse biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Luminex 200, by Merck Group (1 mentions)
Bio plex magpix multiplex reader, by Bio-Rad (1 mentions)
Bio plex pro multiplex immunoassays, by Bio-Rad (1 mentions)
Countur xt, by Bayer (1 mentions)
Siglec f pe clone e50 2440, by BD (1 mentions)
Anti cd45 apc clone 30 f11, by BD (1 mentions)
Cd11c fitc clone hl3, by BD (1 mentions)
Milliplex analyst, by Merck Group (1 mentions)
7 protocols using mouse magnetic luminex screening assay
1
Cytokine Profiling in Mouse Sera
2
Cytokine Profiling in Typhimurium-Infected Mice
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Blood was collected by cardiac puncture from IL-10−/− and WT mice infected i.g with 1 × 105 CFU of S. Typhimurium. Total blood collected was mixed with 100 µl of heparin (125 UI/ml), maintained during 15 min at 37°C and centrifuged at 3,000 × g for 15 min. Serum was collected and stored at −80°C until used. Levels of IL-1β, IL-6, IL-10, IFN-γ, IL-12p70, IL-23p19, and TNF-α were measured on a Luminex 200 (Merck Millipore), using a mouse magnetic luminex screening assay (R&D systems), according to manufacturer instructions.
3
Serum Biomarker Profiling in Mice
4
Biochemical Profiling of Starved Mice
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Prior to bio-clinical analyses, mice were starved for 2 h. After blood collection, bio-clinical analyses were performed by colorimetric methods. In particular, glucose, cholesterol, triglycerides, creatinine, total plasma proteins, albumin, and urea were measured through the automatized KeyLab analyzer (BPCBioSed, Italy) using specific assay kits (BPCBioSed).
Serum adiponectin, FGF21, and leptin levels were measured through a Mouse Magnetic Luminex Screening Assay (R&D System, Minneapolis, MN, USA).
For the glucose tolerance test (OGTT), mice were subjected to fasting for 12 h, followed by oral gavage with 2 g of dextrose/kg body mass. At the indicated time points, blood was collected from the tail vein and glycaemia measured using a glucometer (Bayer Countur XT, Bayer Leverkusen, Germany).
Serum adiponectin, FGF21, and leptin levels were measured through a Mouse Magnetic Luminex Screening Assay (R&D System, Minneapolis, MN, USA).
For the glucose tolerance test (OGTT), mice were subjected to fasting for 12 h, followed by oral gavage with 2 g of dextrose/kg body mass. At the indicated time points, blood was collected from the tail vein and glycaemia measured using a glucometer (Bayer Countur XT, Bayer Leverkusen, Germany).
5
Quantifying Mouse Serum Cytokines
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Mouse cytokines and chemokines in the serum were analyzed using the Mouse Magnetic Luminex Screening Assay (R&D Systems) or the Mouse IL-6 ELISA Kit (MilliporeSigma or RayBiotech).
6
RSV-Induced Lung Inflammation Analysis
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Four days after challenge with RSV A2, mice were sacrificed, and adult and neonate tracheas were cannulated and washed with 700 and 300 μL of PBS, respectively. After BAL fluid was centrifuged, supernatant was stored at -80°C until analysis. Cells were incubated with violet fluorescent live-dead discriminator (Invitrogen, Eugene, OR) for 10 min at room temperature and then washed with 1 mL of PBS and blocked for 5 min with purified CD16/CD32 Fc (clone 2.4G2; BD Pharmingen, San Jose, CA). After blocking, 50 μL of antibody cocktail containing anti-CD45-APC (clone 30-F11), CD11c-FITC (clone HL3), Ly-6G (Gr-1)-PE-Cy7 (clone: 1A8), and Siglec F-PE (clone E50-2440; all from BD Pharmingen) were added to cells and incubated at 4°C for 30 min. Cells were subsequently washed two times with PBS (2% FBS) and fixed with 200 μL of paraformaldehyde. Cells were analyzed by using a BD FACS LSR II flow cytometer and data were analyzed with FlowJo software (version 10; Tree Star, Ashland, OR). The cytokines in BAL supernatant was measured using the mouse Th1/Th2/Th17 BD Cytometric Bead Assay Kit (BD Biosciences, San Jose, CA) according to the manufacturer’s recommendations. The cytokine levels were also analyzed using the Mouse Magnetic Luminex Screening Assay (R&D, Minneapolis, MN) [26 (link)].
7
Cytokine Profiling in LPS-Stimulated Mice
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Blood samples from LPS-stimulated mice were collected from the saphenous vein (2 h after stimulation) and plasma stored at ࣘ-20°C until use. Before cytokine measurements samples were diluted 1:3 and cytokine levels analyzed using the mouse Magnetic Luminex Screening Assay (R&D) custom made for determining TNF-α, IL-2p70, IL-1β, IL-2, IL-4, IL-5, IL-6, IL-10, IL-17A, INF-γ and IL-23p19, GM-CSF levels according to the manufacturer's manual. The standard curves were set and used as recommended, and samples were analyzed with a Milliplex Analyst (Merck Millipore).
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