Biological parameter concentrations of insulin, glucose, adiponectin, ferritin, high-sensitivity C-reactive protein (hs-CRP), interleukin-6 (IL-6), and tumor necrosis factor-alpha (TNFα) were quantified at Hôpital Laval. IL-6, TNFα, insulin, adiponectin and hs-CRP) were measured with Milliplex kits (Millipore, Billerica, MA, USA) in participants’ plasma by Luminex reader (Bio-Rad Lab, Hercules, CA, USA). Glucose concentration was analyzed by Amplex-Red Glucose assay kit, according to the manufacturer's instructions (Life Technologies).
Luminex reader
Manufactured by Bio-Rad
Sourced in United States, United Kingdom
The Luminex reader is a multiplex assay system that utilizes bead-based technology to detect and quantify multiple analytes simultaneously in a single sample. The core function of the Luminex reader is to analyze and measure the fluorescent signals emitted by the beads, providing a highly sensitive and efficient way to perform various immunoassays, protein and nucleic acid detection, and other bioanalytical applications.
Automatically generated - may contain errors
6 protocols using luminex reader
1
Biomarker Quantification in Plasma
2
Multiplex Cytokine Quantification
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cytokines were measured using Bio-Plex assay kits (Bio-Rad) following the manufacturer’s instructions. Briefly, 50 µl of a 1× mixture of magnetic beads coupled with capture Abs for capturing desired cytokines were added to separate wells of flat bottom 96-well microtiter plate. Beads were washed (2×) with Bio-Plex wash buffer. Unless stated otherwise, all incubations were done at room temperature with plates sealed with a plastic sealer and covered with aluminum foil with gentle shaking. All washing was done with Bio-Plex washing buffer. Fifty microliters of diluted standards, blank, or samples was added to individual wells. Plates were incubated for 30 min and washed 3× with Bio-Plex washing buffer. Then 25 µl of 1× mixture of biotinylated detection Abs were added to each well and incubated for 30 min. Plates were washed again, and 50 µl of phycoerythrin-coated streptavidin was added to each well and incubated for 10 min. Plates were then rewashed, and 125 µl of assay buffer was added to each well. Plates were read via Luminex reader (Bio-Rad), and the quantity of each cytokine was deduced using a standard curve included in the assay.
3
Cytokine Profiling of RSV F-Specific Splenocytes
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For cytokine restimulation assays, splenocytes were incubated in 96 well plates with either medium alone (cRPMI-5) or with the pair of RSV F derived MHC II (I-Ed)-binding peptides GWYTSVITIELSNIKE and VSVLTSKVLDLKNYI [9 (link)] (5 μg/mL each) for 72 hours. Supernatants were clarified by centrifugation and stored at-80°C until evaluated. Mouse cytokine/chemokine multiplex kits designed to include IFNγ, IL-5, IL-13, IL-17 and eotaxin (Millipore, Billerica, MA) were used to evaluate restimulated splenocyte supernatants and fresh lung homogenates. Lung homogenates were clarified by centrifugation prior to use. Assays were performed following manufacturer instructions and plates were analyzed on a Luminex reader (Bio-Rad, Hercules, CA). F-specific splenic cytokine production was determined by subtracting media alone values from F stimulated values.
4
Plasma Metabolite Profiling by LC-QTOF
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Plasma concentrations of insulin and adiponectin were determined using Milliplex kits (Millipore, Billerica, MA) and a Luminex reader (Bio-Rad Lab, Hercules, CA). Glucose concentration was measured with the Amplex-Red Glucose assay kit accordingly to the manufacturer’s instructions (Life Technologies).
Plasma concentrations of AA [arginine (Arg), glutamic acid (Glu), isoleucine (Ile), leucine (Leu), methionine (Met), phenylalanine (Phe), tyrosine (Tyr), valine (Val)] and AC [carnitine (C0), acetyl- (C2), propionyl- (C3), butyryl- (C4), isobutyryl- (IsoC4), glutaryl- (C5-DC), hexanoyl- (C6) and octanoylcarnitine (C8)] were determined by isotope-dilution liquid chromatography/hybrid quadrupole-time-of-flight (QTof) mass spectrometry using the method described by Roy et al. [35 ]. Inter-day coefficients of variation ranged from 5.2% to 10.9% for AA and 5.0% to 9.8% for AC. Accuracy ranged from −7.3% to 12.8% for amino acids as assessed against SRM 1950 certified values (National Institute of Standards and Technology, Gaithersburg, MD, USA). We substituted plasma metabolite concentrations below the limit of detection (LOD) by a value equal to half the LOD.
Plasma concentrations of AA [arginine (Arg), glutamic acid (Glu), isoleucine (Ile), leucine (Leu), methionine (Met), phenylalanine (Phe), tyrosine (Tyr), valine (Val)] and AC [carnitine (C0), acetyl- (C2), propionyl- (C3), butyryl- (C4), isobutyryl- (IsoC4), glutaryl- (C5-DC), hexanoyl- (C6) and octanoylcarnitine (C8)] were determined by isotope-dilution liquid chromatography/hybrid quadrupole-time-of-flight (QTof) mass spectrometry using the method described by Roy et al. [35 ]. Inter-day coefficients of variation ranged from 5.2% to 10.9% for AA and 5.0% to 9.8% for AC. Accuracy ranged from −7.3% to 12.8% for amino acids as assessed against SRM 1950 certified values (National Institute of Standards and Technology, Gaithersburg, MD, USA). We substituted plasma metabolite concentrations below the limit of detection (LOD) by a value equal to half the LOD.
5
Multiplex Cytokine Quantification
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cytokines were measured using Bio-Plex assay kits (Bio-Rad, Hercules, CA, USA) following the manufacturer’s instructions. Briefly, 50 μL of a 1× mixture of magnetic beads coupled with capture Abs for capturing desired cytokines were added to separate wells of flat-bottom 96-well micro-titer plate. Beads were washed 2× with Bio-Plex wash buffer. Unless stated otherwise, all incubations were performed at RT with plates sealed with a plastic sealer and covered with aluminum foil with gentle shaking. All washing was performed with Bio-Plex washing buffer. A total of 50 μL of diluted standards, blank, or samples were added to individual wells. Plates were incubated for 30 min and washed 3× with Bio-Plex washing buffer. Then, 25 μL of 1× mixture of biotinylated detection Abs were added to each well and incubated for 30 min. Plates were washed again and 50 μL of phycoerythrin-coated streptavidin was added to each well and incubated for 10 min. Plates were then rewashed and 125 μL of assay buffer was added to each well. Plates were read via Luminex reader (Bio-Rad) and the quantity of each cytokine was deduced using a standard curve included in the assay.
6
Serum IP-10 Quantification Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Blood samples were taken by vena puncture and serum aliquots were stored at À80 C. IP-10 levels were measured with suspension bead assays (Bio-Rad, Richmond, CA) using a high sensitivity Luminex reader (BioRad, BioSource, Linco, Colchester, UK) according to the manufacturer's recommendations. The serum concentration was expressed as pg/ml.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!