The concentration of various biological markers for inflammation and stress in blood and saliva [33 (link), 34 (link)] will be measured using multiplex immunoassay technology (Meso Scale Discovery, MSD). This multi-array technology enables the detection of up to 72 substances in multiplex format. Untargeted biomarker analysis will be performed using proteomics. This will be done by using mass spectrometry in combination with various separation methods such as two-dimensional gel electrophoresis (2-DE) and liquid chromatography (LC). A venous blood sample of about 10 ml is taken from the arm. The saliva sample is taken 15 min after washing the mouth with water by placing a cotton swab (Salivette) in the mouth for 3 min. All samples will be unidentified (marked with a code number). 2-DE instruments in combination with digitizing camera and special software (PDQuest, Bio Rad) for protein separation and quantification are available at the PAINOMICS® laboratory (Linköping University). The laboratory is also equipped with a MESO QUICKPLEX SQ 120 instrument (Meso Scale Discovery, Maryland, USA). EASY-nLC II (Thermo Scientific) combined with LTQ Orbitrap Velos Pro hybrid mass spectrometer (Thermo Scientific) with a nano-electrospray source available at the Core Facility at the Medical Faculty, Linköping University will be used.
Meso quickplex sq 120 instrument
Manufactured by Mesoscale
Sourced in United States
The MESO QUICKPLEX SQ 120 is a multiplex immunoassay instrument designed for quantitative analysis of multiple analytes in a single sample. The instrument utilizes Mesoscale's proprietary electrochemiluminescence (ECL) technology to simultaneously measure a variety of analytes, including proteins, peptides, and small molecules. The MESO QUICKPLEX SQ 120 is capable of processing up to 96 samples per run and can analyze up to 10 different analytes per sample.
Automatically generated - may contain errors
Lab products found in correlation
Discovery workbench data analysis software, by Mesoscale (3 mentions)
Msd gold small spot streptavidin 96 well plates, by Mesoscale (2 mentions)
Ltq orbitrap velos pro hybrid mass spectrometer, by Thermo Fisher Scientific (1 mentions)
Pdquest, by Bio-Rad (1 mentions)
Meso scale discovery (msd), by Mesoscale (1 mentions)
Easy nlc 2, by Thermo Fisher Scientific (1 mentions)
Dpp4 010, by Merck Group (1 mentions)
Rmhmag 84k, by Merck Group (1 mentions)
P2714 1btl, by Merck Group (1 mentions)
U plex assay, by Mesoscale (1 mentions)
Protease inhibitor, by Merck Group (1 mentions)
Mtesr plus, by Merck Group (1 mentions)
Phosphoramidon, by Merck Group (1 mentions)
V plex aβ peptide panel 1 6e10 kit, by Mesoscale (1 mentions)
Msd technology, by Mesoscale (1 mentions)
V plex proinflammatory panel 1 mouse kit, by Mesoscale (1 mentions)
2 d quant kit, by GE Healthcare (1 mentions)
V plex human biomarkers, by Mesoscale (1 mentions)
V plex proinflammatory panel 1 human kit, by Mesoscale (1 mentions)
10 protocols using meso quickplex sq 120 instrument
1
Biomarker Analysis in Blood and Saliva
2
Multiplex Assay for Rat Metabolic Hormones
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Blood samples were collected at termination in EDTA-coated tubes containing DDP-IV inhibitor (DPP4-010, Millipore, Denmark) and protease inhibitor cocktail (P2714-1BTL, Sigma-Aldrich, Denmark). Plasma was separated and stored at −80 °C. Leptin, C-peptide, peptide YY (PYY) and ghrelin were determined in a rat multiplex assay (#RMHMAG-84K, Merck Millipore, Darmstadt, Germany) on a Luminex Flexmap 3D machine (Luminex, Austin, TX). Total and active GLP-1 was determined using single-plex assays (K150JVC/K150JWC, Meso Scale Discovery, Rockville, MD) on a MESO QuickPlex SQ 120 instrument (Meso Scale Discovery, Rockville, MD).
3
Multiplex Immunoassay for Cytokine Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The analysis of inflammatory markers has been described previously.25 (link) In brief, a U-PLEX assay based on an electrochemiluminescent detection method (Meso Scale Diagnostics, Rockville, MD) was used to analyze the concentrations of 71 cytokines in plasma samples from patients (n = 13) with peripheral NeuP and healthy controls (n = 13). A MESO QUICKPLEX SQ 120 instrument equipped with DISCOVERY WORKBENCH data analysis software (Meso Scale Diagnostics, Rockville, MD) was used to collect and analyze data. Electrochemiluminescent signals from calibrators were fitted to a weighted 4-parametric logistic model to form standard curves.
4
DYRK1A Protein Quantification in Plasma
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Immunoassay plates were obtained by spotting biotinylated AC4 (from a set of seven monoclonal antibodies raised against a short form of DYRK1A, 1‐502 aa) on MSD GOLD Small Spot Streptavidin 96‐well plates (Meso Scale Diagnostics, Rockville, MD , USA). After incubation with plasma samples or calibrator samples (serial dilution of DYRK1A protein) MSD GOLD SULFO‐TAG conjugated detection antibody (AC6) was used to quantify DYRK1A protein levels on a MESO QuickPlex SQ120 instrument (Meso Scale Diagnostics) using electrochemiluminescence detection. Plasma samples underwent a single freeze‐thaw cycle before analyses, and all samples were analyzed in duplicate with a coefficient of variability acceptance criteria of <20%.
5
Quantification of Aβ Peptides from hiPSC Conditioned Media
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
hiPSCs were plated on matrigel pre-coated 12-well plates in triplicate. Two days later, medium was replaced by 500µL of mTeSR Plus supplemented with phosphoramidon (10 µM, Sigma-Aldrich). Twenty-four hours later, conditioned medium was collected in each well and centrifuged at 1000×g for 5 min. The supernatant was supplemented with a cocktail of protease inhibitors (Sigma-Aldrich) and frozen at − 80 °C until use. The quantification of Aβ peptides was performed using the MSD technology (M eso S cale D iagnostics, Rockville, MD, USA) with the V-PLEX Aβ Peptide Panel 1 (6E10) kit according to manufacturer’s instructions. Conditioned media were diluted 1:2 in Diluent 35 and analyzed in duplicate. The plate was analyzed using a MESO QuickPlex SQ 120 instrument (Meso Scale Diagnostics). Aβ quantities were normalized to the total protein amount measured in each sample.
6
Cytokine and Intestinal Barrier Assessment
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Serum cytokines, including IL-10, IFN-γ, IL-1β, IL-12p70, IL-2, IL-4, IL-6, IL-5, growth regulates oncogenes (KC/GRO) and TNF-α, were measured using V-PLEX proinflammatory Panel 1 (mouse) kit (Meso Scale Discovery) by a Meso QuickPlex SQ 120 instrument (Meso Scale Discovery) follow the instruction. The mRNA relative expression of TNF-α, IL-10 and IL-1β in jejunum of mice were determined using RT-qPCR. The gene for β-actin was used as an endogenous reference. The relative expression of genes was expressed using the 2−ΔΔCt (Yu et al., 2020 (link)). The primers are listed in Table S1
Concentrations of D-lactate, diamine oxidase (DAO) and immunoglobulins in serum and sIgA in jejunal tissues were determined using corresponding commercially available mouse ELISA kits (Beijing Yonghui Biotechnology Co., Ltd., China) according to procedures described by manufacturers.
Concentrations of D-lactate, diamine oxidase (DAO) and immunoglobulins in serum and sIgA in jejunal tissues were determined using corresponding commercially available mouse ELISA kits (Beijing Yonghui Biotechnology Co., Ltd., China) according to procedures described by manufacturers.
7
Multiplex Cytokine Analysis Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The concentrations of 71 cytokines, chemokines, and growth factors were analyzed using a U-PLEX assay based on an electrochemiluminescent detection method (Meso Scale Diagnostics, Rockville, MD) according to the manufacturer's recommendations. Data were collected and analyzed using MESO QUICKPLEX SQ 120 instrument equipped with DISCOVERY WORKBENCH data analysis software (Meso Scale Diagnostics). Samples were thawed on the day of analysis, blinded, and were randomly mixed.
8
Salivary Inflammatory Protein Profiling
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The concentrations of inflammatory proteins in saliva samples were analyzed from the last six healthy control subjects from the original group of 16 controls with the purpose to investigate the possibility to detect these proteins in the samples. The samples were analyzed using a commercially available V-PLEX human biomarkers (Meso Scale Diagnostics, Rockville, MD, USA) multiplex assay. The assay contained 39 biomarkers (see Table 4) and was based on an electrochemiluminescent detection method and performed exactly according to the manufacturer’s recommendations. Data were collected and analyzed using MESO QUICKPLEX SQ 120 instrument equipped with DISCOVERY WORKBENCH® data analysis software (Meso Scale Diagnostics, Rockville, MD, USA).
The precisions based on both intra- and interassay coefficient of variations were <10% within the detection limits. The detection limit for each investigated biomarker is shown in Table 4 [24 (link),25 (link)].
Total protein concentration was measured using 2D Quant-kit (GE Healthcare Biosciences AB, Uppsala, Sweden), according to the manufacturer’s protocol.
The precisions based on both intra- and interassay coefficient of variations were <10% within the detection limits. The detection limit for each investigated biomarker is shown in Table 4 [24 (link),25 (link)].
Total protein concentration was measured using 2D Quant-kit (GE Healthcare Biosciences AB, Uppsala, Sweden), according to the manufacturer’s protocol.
9
Cytokine Production Profiling of T Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For cytokine production analysis, 0.1 × 106 T cells were cocultured with 5 × 104 target cells (effector/target ratio = 2:1) for 24 hours with or without rapamycin (1 nM, unless otherwise specified) in TCGM. Culture supernatants were evaluated using the V-PLEX Proinflammatory Panel 1 Human kit (Meso Scale Diagnostics) and analyzed by the MESO QuickPlex SQ 120 Instrument (Meso Scale Diagnostics) according to the manufacturer’s instructions.
10
Quantifying DYRK1A Protein Levels
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Immunoassay plates were spotted with biotinylated AC4 (from a set of seven monoclonal antibodies raised against a short form of DYRK1A, 1–502 aa) on MSD GOLD Small Spot Streptavidin 96-well plates (Meso Scale Diagnostics, Rockville, MD, USA). Spotting was performed by the Meso Scale Diagnostics spotting facility. After incubation with plasma samples or calibrator samples (serial dilution of DYRK1A), MSD GOLD SULFO-TAG conjugated detection antibody (AC6) was used to quantify DYRK1A protein levels on a MESO QuickPlex SQ120 instrument (Meso Scale Diagnostics) using electrochemiluminescence detection. AC4 and AC6 were already used in a previous study (9) Plasma samples underwent a single freeze–thaw cycle before analyses. All samples were measured in duplicate with a coefficient of variability acceptance criteria of <20%, and within one round of experiments with the same batch of precoated plates. Baseline and longitudinal samples obtained from each participant were measured side by side in the same run to avoid the effect of run-to-run variability. All analyses were performed by one technician, who was blind to clinical diagnosis. Concentrations are shown in ng/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!