The plasma levels of interferon-inducible protein-10 (IP10)(also known as CXCL10), monocyte chemoattractant protein-1 (MCP-1) (also known as CCL2), and macrophage inflammatory protein-1β (MIP-1β) (also known as CCL4) were measured using a Millipore cytokine three-plex panel assay (MILLIPLEX MAP Human Cytokine/Chemokine Magnetic Bead Panel)(Milliplex MAP kits, EMD Millipore, Billerica, MA, USA). The plasma levels of interferon gamma (IFNγ), tumor necrosis factor-α (TNF-α), interleukin-1β(IL-1β), IL-6, IL-8 (also known as CXCL8), and IL-17A were determined using a Millipore cytokine seven-plex panel assay (MILLIPLEX MAP Human Cytokine/Chemokine Magnetic Bead Panel) (Milliplex MAP kits, EMD Millipore, Billerica, MA, USA). All analyses were performed by T.Y. Chen according to the manufacturer’s protocol. The data were read using a Luminex 200 system (Luminex, Austin, TX, USA). Values of these cytokines and chemokines were reported as pg/ml. Data on cytokines and chemokines were collected and analyzed using an instrument equipped with MILLIPLEX Analyst software (EMD Millipore). For these nine cytokines/chemokines, the intra-assay laboratory coefficients of variation were less than 8% and the inter-assay coefficients of variation were less than 10%.
Milliplex analyst software
Manufactured by Merck Group
Sourced in United States, Italy, Germany
The Milliplex Analyst software is a data analysis tool designed for use with Millipore's Milliplex assay kits. The software provides functionality for data acquisition, analysis, and visualization of multiplex assay results.
Automatically generated - may contain errors
Lab products found in correlation
Milliplex map human cytokine chemokine magnetic bead panel, by Merck Group (7 mentions)
Milliplex map kit, by Merck Group (6 mentions)
Magpix instrument, by Merck Group (4 mentions)
Xponent software, by Merck Group (3 mentions)
Milliplex map kit magnetic bead panels, by Merck Group (3 mentions)
Magpix reader, by Merck Group (3 mentions)
Microplate spectrophotometer, by Bio-Rad (3 mentions)
Magpix milliplex system, by Merck Group (2 mentions)
Milliplex analyzer, by Merck Group (2 mentions)
Mouse cytokine 32 plex kit, by Merck Group (2 mentions)
Rat cytokine chemokine magnetic bead panel kit, by Merck Group (2 mentions)
Neopterin competitive enzyme immunoassay, by ALPCO (2 mentions)
Milliplex magnetic bead panel kits, by Thermo Fisher Scientific (2 mentions)
Milliplex multi analyte panel kit ccytomag 90k, by Merck Group (2 mentions)
Procartaplex panels, by Thermo Fisher Scientific (1 mentions)
Cxcl2, by R&D Systems (1 mentions)
Interleukin 6 (il 6), by R&D Systems (1 mentions)
Ccl11, by R&D Systems (1 mentions)
Cxcl1, by R&D Systems (1 mentions)
Milliplex map human cardiovascular disease magnetic bead panel 1, by Merck Group (1 mentions)
80 protocols using milliplex analyst software
1
Multiplex Cytokine/Chemokine Profiling
2
Multiplex Cytokine Serum Analysis
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Serum concentrations of IL-6, IL-8, IL-10, and TNF-α were measured according to our previously described multiplex technique [17] . Briefly, aliquots of serum were analyzed in duplicate using a high-sensitivity bead-based multiplex assay, according to the manufacturer's recommendations (Milliplex; EMD Millipore; St. Louis, MO). Acquired raw data files were used to determine unknown concentrations using Milliplex Analyst Software (EMD Millipore). Interassay CV was < 9%.
3
Quantifying Immune Responses to Imprime and LPS
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At various times following Imprime treatment or lipopolysaccharide (LPS) injection (i.v., 2 µg), serum, spleens, and sdLNs (pooled inguinal, axillary, and brachial lymph nodes) were harvested and sdLNs were appropriately weighed. Tissues were then manually disrupted, and particulate cellular debris was pelleted in a refrigerated table-top centrifuge at maximum speed. The clarified supernatant was collected and stored at -80°C until analysis. Cytokine and chemokine proteins were detected using mouse pre-designed ProcartaPlex panels (Thermo Fisher Scientific) according to the manufacturer’s instructions. Samples were run on a Luminex xMAP 200 using Luminex xPonent 3.1 software and analyzed using Milliplex Analyst software (Merck Millipore). Data were adjusted for dilution factors and normalized per 10 mg tissue.
4
Multiplex Cytokine Profiling in Mice
5
Quantifying Urinary FABP3 Levels
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Mid-stream urine samples were collected, aliquoted, and stored at −80°C prior to analysis. Urine samples were thawed on ice prior to analysis. To determine the concentration of FABP3 levels in the urine, samples were examined in duplicate using the MILLIPLEX MAP Human Cardiovascular Disease Magnetic Bead Panel 1 (EMD-Millipore; Billerica, MA) (22 ). To minimize any inter-assay variability, all analyses were carried out on the same day. Sample intra-assay and inter-assay coefficients of variability were <10%, which meets the threshold for statistical acceptability (23 (link)). Prior to any sample analysis, Fluidics Verification and Calibration bead kits (Luminex Corp) (24 ) were used to calibrate the MagPix analyzer (Luminex Corp; Austin, Texas) (25 ). At least 50 beads for uFABP3 were acquired using Luminex xPonent software and analyzed using Milliplex Analyst software (v5.1; EMD-Millipore) (26 ).
6
Quantifying Lung Cytokines
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Lungs were instilled with 1 ml HBSS (Corning) containing 0.02 mM EDTA (Lonza) and SIGMAFAST™ Protease Inhibitor Cocktail Tablets (Sigma). Total protein levels of cytokines IL−4, IL−5, and IL−13 in lavage were quantified by mouse–specific Milliplex® multi–analyte kits (EMD Millipore) using a MagPix® instrument platform and related xPONENT® software (Luminex Corporation). The readouts were analyzed with the standard version of the Milliplex Analyst software (EMD Millipore).
7
Cardiovascular Disease Biomarkers Quantification
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Plasma samples were analyzed in duplicate using MILLIPLEX MAP Human Cardiovascular Disease (CVD) Magnetic Bead Panel 1 (EMD-Millipore; Billerica, Mass) to determine the concentrations of FABP3. All sample analyses were completed on the same day to eliminate inter-assay variability. Sample intra-assay and inter-assay coefficient of variation were both less than 10%. The MagPix analyzer (Luminex Corp; Austin, Tex) was calibrated prior to analysis using Fluidics Verification and Calibration bead kits (Luminex Corp). A minimum of 50 beads for each targeted biomarker were acquired using Luminex xPonent software and analyzed using Milliplex Analyst software (v.5.1; EMD-Millipore).
8
Multiplex Cytokine Quantification in Plasma
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Levels of cytokines including IL-6, IL-8, IL-15, MCP-1, TNF-α, G-CSF, GM-CSF, IL-10, IL-1ra, VEGF and IFN-α2 were determined in 25 μl of plasma using a custom-made multiplexed bead-based immunoassay (Milliplex MAP Human Cytokine/Chemokine Magnetic Bead Panel; Merck Millipore, Billerica, MA) in a Luminex 100 Bioanalyzer (Luminex Corp., Austin, TX). Briefly, 25 μL of plasma were added to each well before the addition of 25 μL of premixed microbeads. The plate was incubated overnight at 4 °C with shaking, then washed and reincubated with 25 μL of detection antibody for 1 hour. The plate was washed again and incubated with 25 μL of streptavidin-phycoerythrin for 30 minutes. Finally, after 2 washes the beads were resuspended with 100 μL of sheath fluid and analyzed in the Luminex 100 system. The readouts were analyzed with the standard version of Milliplex Analyst software (Merck Millipore). A five-parameter logistic regression model was used to create standards curves (pg/ml) and to calculate the concentration of each sample.
9
Measuring Plasma Cytokine Levels
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We measured the plasma levels of TNF-α, monocyte chemoattractant protein-1 (MCP-1) (also known as CCL2), and interferon-inducible protein-10 (IP10) (also known as CXCL10) using a Millipore Cytokine Three-Plex Panel Assay (MILLIPLEX MAP Human Cytokine/Chemokine Magnetic Bead Panel) (MILLIPLEX MAP kits, EMD Millipore, Billerica, MA, USA). All analyses were performed in accordance with the manufacturer’s protocol. The data were read using a Luminex 200 system (Luminex, Austin, TX, USA). Data on cytokines and chemokines were collected and analyzed using an instrument equipped with MILLIPLEX Analyst software (EMD Millipore). The intra- and interassay laboratory coefficients of variation were less than 8% and 10%, respectively.
10
Luminex Cytokine Profiling in HTLV-1 and Strongyloidiasis
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Luminex cytokine assays was performed following the manufacturer’s directions on serum samples isolated prior to infection and at the terminal bleed [Fig 1 ]. Briefly, undiluted serum samples isolated from humanized mice infected with HTLV-1, S. stercoralis, or both HTLV-1 and S. stercoralis were analyzed using Milliplex Map Kit magnetic bead panels as per the manufacturer’s protocol (EMDMillipore). Plates were analyzed on a MAGPIX Luminex machine (Austin, TX, USA). All analyte concentrations were calculated using Milliplex Analyst software (EMDMillipore). The following 8 human cytokines were measured in the serum of the control and infected animals: IL-4, IL-5, IL-10, IL-12p40, IL-13, IL-17A, IFN-γ and TNF-α.
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