A total of five blood samples were collected over each 3¼ h meal session, as shown in Figure 1 . Blood glucose (average of triplicates) concentrations (all time points) were measured in serum samples using an auto-calibrating automated sampler (YSI 2300 Stat Plus Glucose L-Lactate analyzer, YSI Inc. Life Sciences, Yellow Springs, OH, USA). Levels of satiety and diabetes-related markers and cytokines (baseline, 1 h post meal, and 2 h post meal) were quantified using multiplex immunoassays (Bio-Plex Pro human diabetes 10-plex immunoassay (# 171A7001M) (measuring C-peptide, ghrelin, GIP, glucagon-like peptide 1 (GLP-1), glucagon, insulin, leptin, total plasminogen activator inhibitor-1 (PAI-1), resistin, and visfatin), Bio-Plex Precision Pro human cytokine 10-plex immunoassay (# 171A1001P) (measuring IL-1β, IL-2, IL-4, IL-5, IL-6, IL-10, IL-12 (p70), IL-13, interferon gamma (IFN-γ) and tumor necrosis factor alpha (TNF-α)) and Bio-Plex Pro human adiponectin single-plex immunoassay (# 171A7003M)) and a Bio-Plex© 200 multiplex suspension array system (Bio-Rad Laboratories, Inc., Hercules, CA, USA), according to the manufacturer’s instructions.
Bio plex 200 multiplex suspension array system
Manufactured by Bio-Rad
Sourced in United States
The Bio-Plex 200 multiplex suspension array system is a laboratory instrument designed for the analysis of multiple analytes in a single sample. The system utilizes magnetic beads coated with specific capture antibodies to detect and quantify multiple targets simultaneously. The instrument provides the capabilities to perform high-throughput, multi-parameter analysis on a wide range of biomolecules, including proteins, cytokines, and other analytes.
Automatically generated - may contain errors
Lab products found in correlation
Bio plex 5, by Bio-Rad (7 mentions)
Luminex xmap technology, by Bio-Rad (3 mentions)
Procartaplex immunoassay, by Thermo Fisher Scientific (1 mentions)
Procartaplex multiplex immunoassay, by Thermo Fisher Scientific (1 mentions)
Bio plex cell lysis kit, by Bio-Rad (1 mentions)
Bio plex pro mouse cytokine 23 plex immunoassay, by Bio-Rad (1 mentions)
11 protocols using bio plex 200 multiplex suspension array system
1
Comprehensive Metabolic Profiling of Post-Prandial Blood Samples
2
Molecular Profiling of Myocardial Inflammation
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qPCR was performed with various primers listed in Table S2 . Multiplex immunoassay was used to determine the concentrations of inflammatory cytokines and chemokines in the myocardium on a Bio-Plex 200 multiplex suspension array system (Bio-Rad). Matrix metalloproteinase zymography was used to examine the activation of matrix metalloproteinases (MMP) 2 and 9 in heart tissue 22 (link). Western blot was used for analysis of protein expression levels of Rac1 and angiogenic factors. Blots were visualized with ChemiDoc Imager and quantified with ImageJ software.
3
Cytokine Profiling in Ovalbumin-Induced Lung Inflammation
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Cytokine concentrations in stored bronchoalveolar lavage fluid supernatants from BALB/c and Δdbl GATA‐1 mice subjected to ovalbumin/aluminium hydroxide were evaluated using the custom ProcartaPlex™ immunoassay (eBioscience, San Diego, CA, USA) according to the manufacturer's specifications. Fluorescent signals were quantified with the Bio‐Plex 200 multiplex suspension array system equipped with Luminex® xMAP® technology combined with the Bio‐Plex 5.0 software (Bio‐Rad, Hercules, CA, USA). All cytokine concentrations were evaluated in duplicates.
4
Multiplex Quantification of Growth Factors
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Concentrations of HGF, SDF-1, FGF2 and PDGF were simultaneously quantified in cell culture supernatant using the ProcartaPlex™ immunoassay (eBioscience, San Diego, CA). Cytokine concentrations were determined using analyte specific capture beads coated with target-specific capture antibodies according to the manufacturer’s specifications. The analytes were detected by biotinylated analyte-specific antibodies. Following binding of the fluorescent detection label, the reporter fluorescent signal was measured with the Bio-Plex 200 multiplex suspension array system and detected with Bio-Plex 5.0 Software (Bio-Rad, Hercules, CA). The sensitivity for the respective cytokines was: HGF: 36,000 pg/ml, SDF-1: 70,000 pg/ml, FGF2: 20,000 pg/ml, PDGF: 30,000 pg/ml. Complete growth medium was used as background control.
5
Cytokine and Corticosterone Quantification
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Cytokine concentrations in plasma samples were evaluated using ProcartaPlexTM immunoassays (eBioscience, San Diego, CA, United States) according to the manufacturer’s specifications (Mayerhofer et al., 2017 (link)). Fluorescent signals were quantified with the Bio-Plex 200 multiplex suspension array system equipped with Luminex® xMAP® technology combined with the Bio-Plex 5.0 software (BioRad, Hercules, CA, United States).
Cytokines that were too low to be detected were excluded from further analysis. All cytokine concentrations were evaluated in duplicates. It should not go unmentioned at this point that IL-1β levels were analyzed in the blood plasma, but were too low to be detected throughout all groups and will not be displayed in the results part.
Corticosterone plasma levels were determined with a specific enzyme immunoassay kit (Assay Designs, Ann Arbor, MI, United States) with a sensitivity of 0.027 ng/ml as previously described (Farzi et al., 2015b (link)) and according to the manufacturer’s specifications.
Cytokines that were too low to be detected were excluded from further analysis. All cytokine concentrations were evaluated in duplicates. It should not go unmentioned at this point that IL-1β levels were analyzed in the blood plasma, but were too low to be detected throughout all groups and will not be displayed in the results part.
Corticosterone plasma levels were determined with a specific enzyme immunoassay kit (Assay Designs, Ann Arbor, MI, United States) with a sensitivity of 0.027 ng/ml as previously described (Farzi et al., 2015b (link)) and according to the manufacturer’s specifications.
6
Multiplex Cytokine Quantification
7
Multiplex Cytokine Quantification in Plasma
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Concentrations of IFN-γ, IL-1β, IL-6 and TNF-α were simultaneously quantified in plasma using the ProcartaPlex™ immunoassay (eBioscience, San Diego, CA, USA). Cytokine concentrations were determined using analyte specific capture beads coated with target-specific capture antibodies according to the manufacturer’s specifications. The analytes were detected by biotinylated analyte-specific antibodies. Following binding of the fluorescent detection label (SA-PE), the reporter fluorescent signal was measured with the Bio-Plex 200 multiplex suspension array system employing Luminex xMAP technology in combination with the Bio-Plex 5.0 Software (Bio-Rad, Hercules, CA).
Standard curves for each analyte were generated by using the reference analyte concentration supplied and concentrations were calculated using a five-parameter logistic curve-fitting method. Cytokines that were not detected were assigned a value of zero. The sensitivity for the respective cytokines was: IFN-γ: 0.09 pg/mL, IL-1β: 0.14 pg/mL, IL-6: 0.21 pg/mL, TNF-α: 0.39 pg/mL.
Plasma samples of experiment 3.3 were run in duplicate. Since the coefficient of variance for the duplicate samples was small, single samples were run subsequently.
Standard curves for each analyte were generated by using the reference analyte concentration supplied and concentrations were calculated using a five-parameter logistic curve-fitting method. Cytokines that were not detected were assigned a value of zero. The sensitivity for the respective cytokines was: IFN-γ: 0.09 pg/mL, IL-1β: 0.14 pg/mL, IL-6: 0.21 pg/mL, TNF-α: 0.39 pg/mL.
Plasma samples of experiment 3.3 were run in duplicate. Since the coefficient of variance for the duplicate samples was small, single samples were run subsequently.
8
Multiplex Cytokine Analysis of T Cell Activation
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Media from T cell activation assays were collected and analyzed using a 9-plex mouse luminex discovery assay targeting RANTES, Granzyme B, IFN-gamma, Il-2, IL-4, IL-6, IL-10, IL-13 and TNF alpha on the Bio-Rad Bio-Plex 200 multiplex suspension array system in the Advanced Analytical Core at UNC Chapel Hill. Media was diluted to 1:5 using assay buffer.
9
Multiplex Immunoassay for Cytokine Quantification
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The plasma levels of IL-6, IL-10, IL-12, and IL-18 were measured with a multiplex immunoassay (ProcartaPlex Multiplex Immunoassays, eBioscience, Vienna, Austria). The assay was performed according to the manufacturer's instructions. The fluorescent signal was measured with the Bio-Plex 200 multiplex suspension array system in combination with the Bio-Plex 5.0 Software (Bio-Rad, Hercules, CA, USA). Standard curves were generated with a five-parameter logistic curve-fitting method. Cytokines that were below detection limit were assigned a value of zero. The sensitivities of the assay were 0.9, 0.35, 0.35, and 8.09 pg/ml for IL-6, IL-10, IL-12, and IL-18, respectively.
10
Multiplex Cytokine Profiling in Heart
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Concentrations of cytokines and chemokines in heart tissue and plasma were measured with the Bio-Plex Pro Mouse Cytokine 23-Plex immunoassay and the Pro TGF-β 3-Plex Immunoassay (Bio-Rad) on a Bio-Plex 200 multiplex suspension array system (Bio-Rad) according to the manufacturer’s protocol. Snap-frozen left ventricles were minced and proteins were isolated with Bio-Plex™ Cell Lysis Kit (Bio-Rad). 1.5 mg of protein per sample was loaded and concentrations of analytes were expressed as pg/mg protein. Undiluted plasma samples were used for multiplex assay and concentrations of analytes were expressed as pg/mL.
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